WO2025137167A2

WO2025137167A2 - Treatment of gpam related diseases and disorders

Info

Publication number: WO2025137167A2
Application number: PCT/US2024/060858
Authority: WO
Inventors: Omri GOTTESMAN; Emma BRANDT; Shannon BRUSE; Brian CAJES; David JAKUBOSKY; David Lewis; Gregory Mcinnes; John VEKICH; David Rozema
Original assignee: Empirico Inc
Current assignee: Empirico Inc
Priority date: 2023-12-20
Filing date: 2024-12-18
Publication date: 2025-06-26
Anticipated expiration: 2026-06-20
Also published as: WO2025137167A3

Abstract

Disclosed herein are compositions comprising an oligonucleotide that targets GPAM. The oligonucleotide may include a small interfering RNA (siRNA) or an antisense oligonucleotide (ASO). Also provided herein are methods of treating conditions associated with GPAM gene mutations that include providing an oligonucleotide that targets GPAM in a subject. Some examples of diseases that may be treated include liver diseases or cardiometabolic diseases.

Description

TREATMENT OF GPAM RELATED DISEASES AND DISORDERS

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63/612,939, filed December 20, 2023, and U.S. Provisional Application No. 63/694,129, filed September 12, 2024, all of which applications are incorporated herein by reference in their entirety.

BACKGROUND

[0002] Metabolic disorders such as liver disorders and cardiovascular disorders are widely abundant, and may affect a wide variety of people. Improved therapeutics are needed for treating these disorders.

SUMMARY

[0003] In certain aspects, disclosed herein is a composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount improves circulating cholesterol, apolipoprotein B, bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase in a subj ect, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69 AS. In some embodiments, the cholesterol comprises total cholesterol, low density lipoprotein cholesterol, or non-high density lipoprotein cholesterol. In some embodiments, the cholesterol is improved by about 10% or more, as compared to prior to administration. In certain aspects, disclosed herein is a composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount improves a liver fibrosis score, non-alcoholic fatty liver disease (NAFLD) activity score, or liver fat percentage in a subject, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61 S, 55S, 56AS, 9 AS, 57AS, 30AS, 58AS, 68AS, and 69AS. In some embodiments, the improvement is by about 10% or more, as compared to prior to administration. In certain aspects, disclosed herein is a composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount decreases a use of statin (HMG Co A reductase inhibitor) medication, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69AS. In some embodiments, the decrease is by about 10% or more, as compared to prior to administration. In certain aspects, disclosed herein is a composition comprising an siRNA that targets GPAM and when administered to a subj ect in an effective amount improves a measurement that reflects a phenotype of esophageal varices, portal hypertension, NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, hepatocellular carcinoma, hyperlipidemia, ischemic heart disease, or coronary heart disease in a subject, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9 AS, 57AS, 30AS, 58AS, 68AS, and 69AS. In some embodiments, the improvementis by about 10% or more, as compared to prior to administration. In certain aspects, disclosed herein is a composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount increases circulating ketone bodies in a subject wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9 AS, 57AS, 30AS, 58AS, 68AS, and 69 AS. In some embodiments, the increase is by about 10% or more, as compared to prior to administration. In some embodiments, the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) moiety, an N-acetylglucosamine (GlcNAc) moiety, or a mannose moiety, attached at a 3’ or 5’ terminus of the oligonucleotide. In some embodiments, the oligonucleotide comprises a GalNAc moiety. In some embodiments, the oligonucleotide further comprises

, wherein

J comprises the oligonucleotide, and wherein J comprises an optional phosphate or phosphorothioate linking to the oligonucleotide. In some embodiments, the siRNA comprises a sense strand and an antisense strand. In some embodiments, the sense strand comprises a sequence comprising at least 19 nucleosides of any one of SEQ ID NO: 1-6354, 13082-13402, 13951-14078, or 14285-14296, or 14337-14339. In some embodiments, the sense strand comprises a sequence comprising at least 15, 16, 17, 18, or 19 consecutive nucleosides of any one of SEQ ID NO: 1-6354, 13082-13402, 13951- 14078, or 14285-14296, or 14337-14339. In some embodiments, the antisense strand comprises a sequence comprising at least 19 nucleosides of any one of SEQ ID NO: 6355-12708, 13403-13723, 14079-14206, 14297-14307, or 14340-14342. In some embodiments, the antisense strand comprises a sequence comprising at least 15, 16, 17, 18, 19 consecutive nucleosides of any one of SEQ ID NO: 6355-12708, 13403-13723, 14079-14206, 14297-14307, or 14340-14342. In certain aspects, disclosed herein is a composition comprising an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 12-30 nucleosides in length, wherein the siRNA comprises sense sequence comprising any one of SEQ ID NO: 13780-13840, 13914-13937, 14254-14255, 14308- 14319, or 14331-14333 or an antisense sequence comprising any one of SEQ ID NO: 13841 -13913, 13938-13950, 14207-14253, 14256-14276, 14277-14284, 14320-14330, or 14334-14336. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, disclosed herein is a method of treating a subject having liver disease, comprising administering an effective amount of the composition disclosed herein to the subject. In some embodiments, the liver disease comprises NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, disclosed herein is a method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition disclosed herein to the subject. In some embodiments, the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease. In certain aspect, disclosed herein is a method of treating a subject having liver disease, comprising administering an effective amount of the composition as described above to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist. In some embodiments, the liver disease comprises NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, dulaglutide, tirzepatide, dulaglutide, semaglutide or a combination thereof. In certain aspects, disclosed herein is a method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition as described above to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist. In some embodiments, the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease In some embodiments, the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, dulaglutide, tirzepatide, dulaglutide, semaglutide or a combination thereof. In certain aspect, disclosed herein is a composition comprising an oligonucleotide that inhibits the expression of GPAM in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 12-30 nucleosides in length, wherein the siRNA comprises sense sequence comprising any one of SEQ ID NO: 13780-13840, 13914-13937, 14254-14255, 14308-14319, or 14331-14333, or an antisense sequence comprising any one of SEQ ID NO: 13841-13913, 13938-13950, 14207-14253, 14256- 14276, 14277-14284, 14320-14330, or 14334-14336. In certain aspects, disclosed herein is a method of treating a subject having liver disease, the method comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, glucose-dependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor- 15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator-activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activin E expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof. In certain aspects, disclosed herein is a method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, glucosedependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor- 15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator- activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activinE expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof.

DETAILED DESCRIPTION

[0004] Large-scale human genetic data can improve the success rate of pharmaceutical discovery and development. A Genome Wide Association Study (GW AS) detects associations between genetic variants and traits in a population sample, and this improves understanding of the biology of disease and provides evidence of applicable treatments. A GWAS generally utilizes genotyping and/or sequencing data, and often involves an evaluation of millions of genetic variants that are relatively evenly distributed across the genome. The most common GWAS design is the case-control study, which involves comparing variant frequencies in cases versus controls. If a variant has a significantly different frequency in cases versus controls, that variant is considered associated with disease. Association statistics used in a GWAS include p-values, as a measure of statistical significance; odds ratios (OR), as a measure of effect size; or beta coefficients (beta), as a measure of effect size. Researchers often assume an additive genetic model and calculate an allelic odds ratio, which is the increased (or decreased) risk of disease conferred by each additional copy of an allele (compared to carrying no copies of that allele). An additional concept in design and interpretation of GW AS is that of linkage disequilibrium, which is the non-random association of alleles. The presence of linkage disequilibrium can obfuscate which variant is “causal.”

[0005] Functional annotation of variants and/or wet lab experimentation is used to identify a causal genetic variant identified via GWAS, and in many cases leads to identification of disease-causing genes. In particular, understanding the functional effect of a causal genetic variant (for example, loss of protein function, gain of protein function, increase in gene expression, or decrease in gene expression) allows that variant to be used as a proxy for therapeutic modulation of the target gene, or to gain insight into potential therapeutic efficacy and safety of a therapeutic that modulates that target. [0006] Identification of such gene-disease associations has provided insights into disease biology and is used to identify novel therapeutic targets for the pharmaceutical industry. In order to translate the therapeutic insights derived from human genetics, disease biology in patients is exogenously ‘programmed’ into replicating the observation from human genetics. There are several options for therapeutic modalities that may be brought to bear in translating therapeutic targets identified via human genetics into novel medicines. These include well established therapeutic modalities such as small molecules and monoclonal antibodies, maturing modalities such as oligonucleotides, and emerging modalities such as gene therapy and gene editing. The choice of therapeutic modality depends on factors such as the location of a target (for example, intracellular, extracellular, or secreted), a relevant tissue (for example, fat or liver) and a relevant indication.

[0007] The GPAM (also known as GPAT or GPAT1) gene is located on chromosome 10, and encodes glycerol-3-phosphate acyltransferase, mitochondrial (GPAM). GPAM may include 828 amino acids and have a mass of about 94 kDa. GPAM may be expressed in liver, adipose, adrenal, thyroid, heart, gall bladder, brain, salivary gland, and testis cells. GPAM may be intracellular. GPAM may exist as two different enzymatic forms, in the mitochondria or in the endoplasmic reticulum. GPAM may catalyze the initial and committing step in glycerolipid biosynthesis and can play a significant role in the regulation of cellular triacylglycerol and phospholipid levels. The mitochondrial enzyme GPAM may preferentially use saturated fatty acids as a substrate for the synthesis of glycerolipids. GPAM may catalyze the first step in this metabolic pathway. GPAM may interact with APP, SREBF1, AGP ATI, AGPAT2, AGPAT3, AGPAT4, AGPAT5, AGPAT6, AGPAT9, GPD1, or MBOAT2. An example of an 828 amino acid sequence, and further description of GPAM is included at uniprot.org under accession no. Q9HCL2 (last modified May 25, 2022). A potential alternatively spliced isoform of GPAM producing a 710 amino acid sequence is included at uniprot.org under accession no. Q5VW52 (last modified Dec. 07, 2004).

[0008] Here it is shown that genetic loss-of-function GPAM variants result in protective blood ketone associations, liver disease-related associations, liver function associations, and blood lipid associations. Therefore, inhibition of GPAM may serve as a therapeutic for treatment of liver or cardiometabolic diseases or disorders such as non-alcoholic fatty liver disease (NAFLD, also known as metabolic dysfunction-associated steatotic liver disease (MASLD)), non-alcoholic steatohepatitis (NASH, also known as metabolic dysfunction-associated steatohepatitis (MASH)), alcoholic liver disease, liver fibrosis, liver cirrhosis, hepatocellular carcinoma, hyperlipidemia, ischemic heart disease, or coronary heart disease. The GPAM inhibition may result in an improved liver function, cardiovascular function, and metabolic phenotypes including favorable liver fat percentage, liver fibrosis score, NAFLD activity score, liver enzyme function test, serum metabolite test, or serum lipid panel test.

[0009] Disclosed herein are compositions comprising an oligonucleotide that targets GPAM. Where inhibition or targeting of GPAM is disclosed, it is contemplated that some embodiments may include inhibiting or targeting a GPAM protein or GPAM RNA. For example, by inhibiting or targeting an RNA (e.g., mRNA) encoded by the GPAM gene using an oligonucleotide described herein, the GPAM protein may be inhibited or targeted as a result of there being less production of the GPAM protein by translation of the GPAM RNA; or a GPAM protein may be targeted or inhibited by an oligonucleotide that binds or interacts with a GPAM RNA and reduces production of the GPAM protein from the GPAM RNA. Thus, targeting GPAM may refer to binding a GPAM RNA and reducing GPAM RNA or protein levels. The oligonucleotide may include a small interfering RNA (siRNA) or an antisense oligonucleotide (ASO). Administration of the oligonucleotide to a subject may improve (e.g., decrease or increase) a liver fat percentage, liver fibrosis score, NAFLD activity score, blood alanine aminotransferase (ALT), blood aspartate aminotransferase (AST), blood alkaline phosphatase (ALP), blood bilirubin, low-density lipoprotein (LDL), total cholesterol, non-HDL cholesterol, or apolipoprotein B (APOB) measurement, or a combination thereof in the subject. Also provided herein are methods of treating a liver or cardiometabolic disorder by providing an oligonucleotide that targets GPAM to a subject in need thereof.

I. COMPOSITIONS

[0010] Disclosed herein, in some embodiments, are compositions comprising an oligonucleotide. In some embodiments, the composition comprises an oligonucleotide that targets GPAM. In some embodiments, the composition consists of an oligonucleotide that targets GPAM. In some embodiments, the oligonucleotide reduces GPAM mRNA expression in the subject. In some embodiments, the oligonucleotide reduces GPAM protein expression in the subject. The oligonucleotide may include a small interfering RNA (siRNA) described herein. The oligonucleotide may include an antisense oligonucleotide (ASO) described herein. In some embodiments, a composition described herein is used in a method of treating a disorder in a subject in need thereof. Some embodiments relate to a composition comprising an oligonucleotide for use in a method of treating a disorder as described herein. Some embodiments relate to use of a composition comprising an oligonucleotide, in a method of treating a disorder as described herein. In some embodiments, the siRNA comprises a modification patern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 157S, 158S, 159S, 160S, 161S, 162S, 163S, 164S, 68AS, 69AS, 70AS, 71AS, and 72AS. In some embodiments, the siRNA comprises sense sequence comprising any one of SEQ ID NO: 13780-13840, 13914-13937, 14254-14255, 14308-14319, or 14331-14333or an antisense sequence comprising any one of SEQ ID NO: 13841-13913, 13938-13950, 14207-14253, 14256- 14276, 14277-14284, 14320-14330, or 14334-14336.

[0011] Some embodiments include a composition comprising an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases GPAM mRNA or protein levels in a cell, fluid or tissue. In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases GPAM mRNA levels in a cell or tissue. In some embodiments, the tissue is liver tissue. In some embodiments, the tissue is fat tissue. In some embodiments, the cell is a hepatocyte. In some embodiments, the cell is an adipocyte. In some embodiments, the GPAM mRNA levels are decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by about 10% or more, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the GPAM mRNA levels are decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages. [0012] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases GPAM protein levels in a cell, fluid or tissue. In some embodiments, the composition decreases GPAM protein levels in a cell or tissue. In some embodiments, the tissue is liver tissue. In some embodiments, the tissue is fat tissue. In some embodiments, the cell is a hepatocyte. In some embodiments, the cell is an adipocyte. In some embodiments, the GPAM protein levels are decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration In some embodiments, the GPAM protan levels are decreased by about 10% or more, as compared to prior to administration. In some embodiments, the GPAM protein levels are decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the GPAM protein levels are decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the GPAM protein levels are decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the GPAM protein levels are decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the GPAM protein levels are decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0013] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount improves a liver disease phenotype. The liver disease may include non-alcoholic fatty liver disease (NAFLD, also known as metabolic dysfunction- associated steatotic liver disease (MASLD)), non-alcoholic steatohepatitis (NASH, also known as metabolic dysfunction-associated steatohepatitis (MASH)), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, the liver disease phenotype is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the liver disease phenotype is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the liver disease phenotype is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the liver disease phenotype is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the liver disease phenotype is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the liver disease phenotype is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the liver disease phenotype is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0014] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount improves a cardiometabolic disease phenotype. The cardiometabolic disease may include hyperlipidemia, ischemic heart disease, or coronary heart disease. In some embodiments, the cardiometabolic disease phenotype is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by about 10% or more, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the cardiometabolic disease phenotype is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages. [0015] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subj ect in an effective amount improves a protective phenotype against a liver disease in the subject. The liver disease may include non-alcoholic fatty liver disease (NAFLD, also known as metabolic dysfunction-associated steatotic liver disease (MASLD)), non-alcoholic steatohepatitis (NASH, also known as metabolic dysfunction-associated steatohepatitis (MASH)), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, the protective phenotype is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 10% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.

[0016] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount improves a protective phenotype against a cardiometabolic disease in the subject. The cardiometabolic disease may include hyperlipidemia, ischemic heart disease, or coronary heart disease. In some embodiments, the protective phenotype is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 10% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration. In some embodiments, the protective phenotype is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.

[0017] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating cholesterol in the subject. The circulating cholesterol may include total cholesterol or non-high density lipoprotein (HDL) cholesterol. The circulating cholesterol may include total cholesterol. The circulating cholesterol may include non-HDL cholesterol. In some embodiments, the circulating cholesterol is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating cholesterol is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating cholesterol is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating cholesterol is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to priorto administration In some embodiments, the circulating cholesterol is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating cholesterol is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating cholesterol is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages. [0018] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating low density lipoproteins (LDL) in the subject. In some embodiments, the circulating LDL is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating LDL is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0019] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating apolipoprotein B (APOB) in the subject. In some embodiments, the circulating APOB is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating APOB is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0020] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating alanine aminotransferase (ALT) in the subject. In some embodiments, the circulating ALT is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating ALT is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0021] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating aspartate aminotransferase (AST) in the subject. In some embodiments, the circulating AST is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating AST is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating AST is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating AST is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7. 5%, as compared to prior to administration. In some embodiments, the circulating AST is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating AST is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating AST is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0022] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating alkaline phosphatase (ALP) in the subject. In some embodiments, the circulating ALP is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating ALP is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0023] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases circulating bilirubin in the subject. In some embodiments, the circulating bilirubin is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the circulating bilirubin is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0024] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount improves a nonalcoholic fatty liver disease (NAFLD) activity score in the subject. In some embodiments, the NAFLD activity score is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by about 10% or more, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the NAFLD activity score is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0025] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases a liver fat percentage in the subject. In some embodiments, the liver fat percentage is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the liver fat percentage is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0026] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subj ect in an effective amount improves a liver fibrosis score in the subj ect. In some embodiments, the liver fibrosis score is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by about 10% or more, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by no more than about 10%, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the liver fibrosis score is improved by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0027] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount decreases a use of an HGM Co A reductase inhibitor (e.g. , a statin) medication in the subject. In some embodiments, the use of statin medications is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by about 10% or more, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by no more than about 10%, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, or no more than about 90%, as compared to prior to administration. In some embodiments, the use of statin medications is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages. [0028] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount increases circulating ketone bodies in the subject. In some embodiments, the circulating ketone body 3 -hydroxy butyrate is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxy butyrate is increased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating ketone body 3- hydroxybutyrate is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by no more than about 2.5%, no more than about 5%, or no more than about 7. 5%, as compared to prior to administration In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration. In some embodiments, the circulating ketone body 3-hydroxybutyrate is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by arange defined by any of the two aforementioned percentages.

[0029] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount increases circulating ketone bodies in the subject. In some embodiments, the circulating ketone body acetoacetate is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration. In some embodiments, the circulating ketone body acetoacetate is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.

[0030] In some embodiments, the composition comprises an oligonucleotide that targets GPAM and when administered to a subject in an effective amount increases circulating ketone bodies in the subject. In some embodiments, the circulating ketone body acetone is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by about 10% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration. In some embodiments, the circulating ketone body acetone is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.

A.siRNAs

[0031] In some embodiments, the composition comprises an oligonucleotide that targets GPAM, wherein the oligonucleotide comprises a small interfering RNA (siRNA). In some embodiments, the composition comprises an oligonucleotide that targets GPAM, wherein the oligonucleotide comprises a small interfering RNA (siRNA) comprising a sense strand and an antisense strand.

[0032] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand is 12-30 nucleosides in length. In some embodiments, the composition comprises a sense strand that is 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers. The sense strand may be 14-30 nucleosides in length. In some embodiments, the composition comprises an antisense strand is 12-30 nucleosides in length. In some embodiments, the composition comprises an antisense strand that is 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers. The antisense strand may be 14-30 nucleosides in length. In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 12-30 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising about 12-30 contiguous nucleosides of a full-length human GPAM mRNA sequence such as SEQ ID NO: 12867. In some embodiments, at least one of the sense strand and the antisense strand comprise a nucleoside sequence comprising at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more contiguous nucleosides of one of SEQ ID NO: 12867. The siRNA may include one or more internucleoside linkages and/or one or more nucleoside modifications. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 12867. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of SEQ ID NO: 12867. Any of the aforementioned siRNAs may include a sense strand of SEQ ID NO: 12867, wherein the nucleotide at the 3’ end of the sense strand sequence has been modified to an A. Any of the aforementioned siRNAs may include an antisense strand of SEQ ID NO: 12867, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. [0033] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand of the composition 5’ -(N)_0-7(N_B)(N)_0-7 -3’ and an antisense strand of composition 5’-(N)_0-7(N_A)(N)_0-7 -3’. Where (N) _0-7 are independently nucleic acid stretches of 0 to 7 nucleotides, (N_B) and (N_A) contain at least 15 contiguous nucleotides from Table 8 and contain no cyclic ribose nucleotides with 2’ hydroxyl groups. The sense and antisense strand have 1-5 phosphorothioate inter -nucleotide linkages and either the sense or antisense strand has a targeting ligand attached to the 5’ or 3’ end. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand composition. The targeting ligand may be a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. Representative examples of the GalNAc moiety is ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6]s[Gal- 6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety is ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL 18, ETL19, ETL 20, ETL21, ETL22 or ETL28. Preferably, the lipid moiety is ETL20. Representative examples of integrin or integrin targeting ligand is epithelial-specific integrin, integrin alpha- v-beta-6 (αvβ6) or integrin alpha-v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide.

[0034] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double -stranded RNA duplex. In some embodiments, the first base pair of the double-stranded RNA duplex is an AU base pair.

[0035] In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides.

[0036] In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides.

[0037] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a 19mer in a human GPAM mRNA. In some embodiments, the siRNA binds with a 12mer, a 13mer, a 14mer, a 15mer, a 16mer, a 17mer, a 18mer, a 19mer, a 20mer, a 21mer, a 22mer, a 23mer, a 24mer, or a 25mer in a human GPAM mRNA. [0038] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a 17mer in a non-human primate GPAM mRNA. In some embodiments, the siRNA binds with a 12mer, a 13mer, a 14mer, a 15mer, a 16mer, a 17mer, a 18mer, a 19mer, a20mer, a21mer, a22mer, a23mer, a24mer, or a 25mer in a non-human primate GPAM mRNA.

[0039] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a human GPAM mRNA and less than or equal to 20 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 10 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 30 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 40 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 50 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 10 human off- targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 20 human off-targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 30 human off-targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 40 human off-targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human GPAM mRNA and less than or equal to 50 human off- targets, with no more than 3 mismatches in the antisense strand. [0040] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, siRNA binds with a human GPAM mRNA target site that does not harbor an SNP, with aminor allele frequency (MAF) greater or equal to 1% (pos. 2-18). In some embodiments, the MAF is greater or equal to about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20%.

[0041] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 1-6354. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 1-6354, at least 80% identical to any one of SEQ ID NOs: 1-6354, at least 85% identical to of any one of SEQ ID NOs: 1-6354, at least 90% identical to any one of SEQ ID NOsl-6354, or at least 95% identical to any one of SEQ ID NOs: 1-6354. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 1-6354, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 1-6354, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 1-6354. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 1-6354. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise an LDL receptor ligand.

[0042] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 6355-12708. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 6355- 12708, at least 80% identical to any one of SEQ ID NOs: 6355-12708, at least 85% identical to of any one of SEQ ID NOs: 6355-12708, at least 90% identical to any one of SEQ ID NOs: 6355-12708, or at least 95% identical to any one of SEQ ID NOs: 6355-12708. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 6355-12708, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 6355-12708, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 6355-12708. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand.

[0043] Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 1-6354. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of SEQ ID NO: 6355-12708. In any of SEQ ID NOs: 1-6354, thymine (T) may be replaced with uracil (U). Any of the aforementioned siRNAs may include an antisense strand where the 5’ nucleoside has been modified to an A. Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U or T. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A, T, C, U, or G. In some embodiments, position 6 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A, T, C, U, or G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A, T, C, U, or G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A. In some embodiments, position 6 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to an A. In some embodiments, position 1 and position 6, position 1 and position 19, position 6 and position 19, or position 1, position 6, and position 19 of any one of SEQ ID NOs: 1-6354 is modified to an A. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a T or U. In some embodiments, position 6 (from the 5’ end) of any one of SEQ ID NOs: 1 -6354 is modified to a T or U. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to aT or U. In some embodiments, position 1 and position 6, position 1 and position 19, position 6 and position 19, or position 1, position 6, and position 19 of any one of SEQ ID NOs: 1-6354 is modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a G. In some embodiments, position 6 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a G. In some embodiments, position 1 and position 6, position 1 and position 19, position 6 and position 19, or position 1, position 6, and position 19 of any one of SEQ ID NOs: 1-6354 is modified to a G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a C. In some embodiments, position 6 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a C. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 1-6354 is modified to a C. In some embodiments, position 1 and position 6, position 1 and position 19, position 6 and position 19, or position 1, position 6, and position 19 of any one of SEQ ID NOs: 1-6354 is modified to a C.

[0044] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 13082-13402. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13082-13402, at least 80% identical to any one of SEQ ID NOs: 13082-13402, at least 85% identical to of any one of SEQ ID NOs: 13082-13402, at least 90% identical to any one of SEQ ID NOs: 13082-13402, or at least 95% identical to any one of SEQ ID NOs: 13082-13402. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13082-13402, or an sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13082-13402, or an sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13082-13402. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5 ’ to 3 ’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein. The sense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NO: 13082-13402. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise a LDL receptor ligand.

[0045] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13403-13723. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13403- 13723, at least 80% identical to any one of SEQ ID NOs: 13403-13723, at least 85% identical to of any one of SEQ ID NOs: 13403-13723, at least 90% identical to any one of SEQ ID NOs: 13403- 13723, or at least 95% identical to any one of SEQ ID NOs: 13403-13723. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13403- 13723, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13403-13723, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13403-13723. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand.

[0046] Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 13082-13402. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of SEQ ID NO: 13403-13723. In any of SEQ ID NOs: 13082-13402, thymine (T) may be replaced with uracil (U). Any of the aforementioned siRNAs may include a sense strand wherein the 3’ nucleoside has been modified to an A. Any one of the aforementioned siRNAs may include a sense strand sequence wherein the 5’ nucleoside has been modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an A, T, C, U, or G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an A, T, C, U, or G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an A, T, C, U, or G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13082-13402 is modified to an A, T, C, U, or G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an A. In some embodiments, position 14 (from the 5’ end) of the sense strand of any one of SEQ ID NOs: 13082- 13402 is modified to an A. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an A. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13082-13402 is modified to an A. In some embodiments, position 1 (from the 5’ end of any one of SEQ ID NOs: 13082-13402 is modified to a T or U. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to a T or U. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to a T or U. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13082-13402 is modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to an G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13082-13402 is modified to an G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to a C. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to a C. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13082-13402 is modified to a C. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13082-13402 is modified to a C.

[0047] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 13951-14078. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13951-14078, at least 80% identical to any one of SEQ ID NOs: 13951-14078, at least 85% identical to of any one of SEQ ID NOs: 13951-14078, at least 90% identical to any one of SEQ ID NOsl3951-1407813951- 14078, or at least 95% identical to any one of SEQ ID NOs: 13951-14078. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13951- 14078, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13951-14078, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13951-14078. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NO: 13951-14078. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise an LDL receptor ligand. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0048] Any of the aforementioned siRNAs may include a sense strand wherein the 3’ nucleoside has been modified to an A. Any one of the aforementioned siRNAs may include a sense strand sequence wherein the 5’ nucleoside has been modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an A, T, C, U, or G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an A, T, C, U, or G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an A, T, C, U, or G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13951-14078 is modified to an A, T, C, U, or G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an A. In some embodiments, position 14 (from the 5’ end) of the sense strand of any one of SEQ ID NOs: 13951-14078 is modified to an A. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an A. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13951-14078 is modified to an A. In some embodiments, position 1 (from the 5’ end of any one of SEQ ID NOs: 13951-14078 is modified to a T or U. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to a T or U. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to a T or U. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13951-14078 is modified to aT or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to an G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13951-14078 is modified to an G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to a C. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to a C. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 13951-14078 is modified to a C. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 13951-14078 is modified to a C.

[0049] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 14079-14206. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 14079- 14206, at least 80% identical to any one of SEQ ID NOs: 14079-14206, at least 85% identical to of any one of SEQ ID NOs: 14079-14206, at least 90% identical to any one of SEQ ID NOs: 14079- 14206, or at least 95% identical to any one of SEQ ID NOs: 14079-14206. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14079- 14206, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14079-14206, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 14079-14206. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 14079-14206. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 13951-14078. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of SEQ ID NO: 14079-14206.

[0050] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 14285-14296 or 14337-14339. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 14285- 14296 or 14337-14339, at least 80% identical to any one of SEQ ID NOs: 14285-14296 or 14337- 14339, at least 85% identical to of any one of SEQ ID NOs: 14285-14296 or 14337-14339, at least 90% identical to any one of SEQ ID NOs: 14285-14296 or 14337-14339, or at least 95% identical to any one of SEQ ID NOs: 14285-14296 or 14337-14339. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14285-14296 or 14337- 14339, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14285-14296 or 14337-14339, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 14285-14296 or 14337-14339. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NO: 14285-14296 or 14337-14339.

[0051] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 14297-14307 or 14340-14342. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 14297-14307 or 14340-14342, at least 80% identical to any one of SEQ ID NOs: 14297- 14307 or 14340-14342, at least 85% identical to of any one of SEQ ID NOs: 14297-14307 or 14340- 14342, at least 90% identical to any one of SEQ ID NOs: 14297-14307 or 14340-14342, or at least 95% identical to any one of SEQ ID NOs: 14297-14307 or 14340-14342. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14297- 14307 or 14340-14342, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14297-14307 or 14340-14342, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 14297-14307 or 14340-14342. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety.

[0052] Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 14285-14296 or 14337-14339. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of SEQ ID NO: 14297-14307 or 14340-14342. In any of SEQ ID NOs: 14285-14296 or 14337-14339, thymine (T) may be replaced with uracil (U). Any of the aforementioned siRNAs may include a sense strand wherein the 3’ nucleoside has been modified to an A. Any one of the aforementioned siRNAs may include a sense strand sequence wherein the 5’ nucleoside has been modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A, T, C, U, or G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A, T, C, U, or G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 14285- 14296 or 14337-14339 is modified to an A, T, C, U, or G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A, T, C, U, or G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A. In some embodiments, position 14 (from the 5’ end) of the sense strand of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337- 14339 is modified to an A. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an A. In some embodiments, position 1 (from the 5’ end of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to a T or U. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337- 14339 is modified to a T or U. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to aTor U. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to a T or U. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337- 14339 is modified to an G. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an G. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an G. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to an G. In some embodiments, position 1 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to aC. In some embodiments, position 14 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to a C. In some embodiments, position 19 (from the 5’ end) of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to a C. In some embodiments, position 1 and position 14, position 1 and position 19, position 14 and position 19, or position 1, position 14, and position 19 of any one of SEQ ID NOs: 14285-14296 or 14337-14339 is modified to a C.

[0053] In some embodiments, any of the aforementioned siRNA comprising a sense stand comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NOS: 1-6354, 13082-13402, 13951-14078, 14285-14296 or 14337-14339. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal- 6]s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28.

Preferably, the lipid moiety is ETL20. Representative examples of integrin or integrin targeting ligand is epithelial-specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha-v-beta-3 or arginine- glycine-aspartic acid (RGD) peptide. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0054] In some embodiments, any of the aforementioned siRNA comprising an antisense stand comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NO: 6355-12708, 13403-13723, 14079-14206, 14297- 14307 or 14340-14342. The antisense stand may comprise a lipid moiety. The antisense strand may comprise a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6] s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28. Preferably, the lipid moiety is ETL20. Representative examples of integrin or integrin targeting ligand is epithelial -specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha-v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0055] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset A. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset A. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset A, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset A, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset A. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset A. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset A, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. [0056] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset B. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset B. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset B, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset B, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset B. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset B. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset B. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset

B, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T.

[0057] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset C. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset C. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset C, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset C, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset C. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset C. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset C. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset

C, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T.

[0058] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset D. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset D. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset D, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset D, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset D. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset D. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset D. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset D, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. [0059] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset E. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset E. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset E, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset E, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset E. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset E. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset E. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset E, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T.

[0060] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset F. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset F. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset F, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset F, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset F. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset F. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset F. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset F, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T.

[0061] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset I. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset I. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset I, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset I, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset I. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset I. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset I. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset I, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset I). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset I.

[0062] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset J. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset J. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset J, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset J, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset J. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset J. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset J. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset J, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset J). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset J.

[0063] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset K. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset K. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset K, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset K, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset K. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset K. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset K. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset K, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T.

[0064] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset L. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset L. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset L, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset L, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset L. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset L. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset L. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset L, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset L). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset L.

[0065] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset M. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset M. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset M, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset M, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset M. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset M. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset M. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset M, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset M). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset M.

[0066] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset N. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset N. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset N, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset N, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset N. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset N. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset N. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset N, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset N). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset N.

[0067] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset O. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset O. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset O, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset O, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset O. The sense strand or antisense strand may comprise any modifications described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of subset O. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of one of subset O. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. Any of the aforementioned siRNAs may include an antisense strand of one of subset O, wherein the nucleotide at the 5’ end of the sense strand sequence has been modified to a T. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset O). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset O.

[0068] In some embodiments, any of the aforementioned siRNA comprising a sense strand or antisense stand comprises a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense or anti sense strand sequence of one of Subset A, subset B, subset C, subset D, subset E, subset F, subset I, subset J, subset K, subset L, subset M, subset N or subset O. The sense strand or antisense stand may comprise a lipid moiety. The antisense strand may comprise a GalNAc moiety, sense strand or antisense stand may comprise an integrin or an integrin targeting ligand. The sense strand or antisense stand may comprise an angiopep-2. The sense strand or antisense stand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The sense strand or antisense stand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6] s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28. Preferably, the lipid moiety is ETL20. Representative examples of integrin or integrin targeting ligand is epithelial -specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha- v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0069] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA herein (such as an siRNA in a table herein). In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of an siRNA herein. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of an siRNA herein, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of an siRNA herein, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of an siRNA herein. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety. In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein. The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 2856-3037.

Representative example of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6]s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative example of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL22, or ETL28. Preferably, the lipid moiety is ETL20. Representative example of integrin or integrin targeting ligand includes, but is not limited to, epithelial -specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha-v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide. [0070] Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A. Any of the aforementioned siRNAs may include a sense strand where the 3’ nucleoside has been modified to an A at position 19 (from the 5’ end). Any one of the aforementioned siRNAs may include an antisense strand sequence wherein the 5’ nucleoside has been modified to a U. In some embodiments, the composition comprises an oligonucleotide that inhibits or reduces the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand comprise a oligonucleotide sequences as disclosed in Table 8, 13, 17, 21, 24, 30, 34, 48, 62,

67. 68, 75, 79, 83, 86, 91, 96, or 101, each strand is independently about 19-21 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising at least about 15 contiguous nucleosides of oligonucleotide sequences as disclosed in Table 8, 13, 17, 21, 24, 30, 34, 48, 62, 67, 68, 75, 79, 83, 86, 91, 96, or 101

[0071] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand comprise a oligonucleotide sequences as disclosed in Tables 8, 13, 17, 21, 24, 27,

30. 34. 48. 62. 64. 65. 67. 68, 75, 79, 83, 86, 91, 96 and 101, each strand is independently about 19- 21 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising at least about 15 contiguous nucleosides of oligonucleotide sequences as disclosed in Table 8, 13, 17, 21, 24, 27, 30, 34, 48, 62, 64, 65, 67, 68, 75, 79, 83, 86, 91, 96 and 101

[0072] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ IDNOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166, 13167, 13168, 13169, 13170,

13171, 13172, 13173, 13100, 13157, 13158, 13159, 13160, 13161, 13162, 13163, 13164, 13344,

13103, 13216, 13217, 13218, 13219, 13220, 13221, 13222, 13223, 13224, 13307, 13308, 13309,

13310, 13345, 13346, 13106, 13267, 13268, 13269, 13270, 13271, 13272, 13273, 13998, 13974,

13980, and 13274. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165,

13166, 13167, 13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158, 13159, 13160,

13161, 13162, 13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220, 13221, 13222,

13223, 13224, 13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268, 13269, 13270,

13271, 13272, 13273, 13998, 13974, 13980, and 13274., at least 80% identical to any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166, 13167, 13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158, 13159, 13160, 13161, 13162, 13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220, 13221, 13222, 13223, 13224, 13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268, 13269, 13270, 13271, 13272, 13273, 13998, 13974, 13980, and 13274., at least 85% identical to of any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166,

13167, 13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158, 13159, 13160, 13161,

13162, 13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220, 13221, 13222, 13223,

13224, 13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268, 13269, 13270, 13271,

13272, 13273, 13998, 13974, 13980, and 13274., at least 90% identical to any one of SEQ ID NOsl3329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166, 13167, 13168, 13169, 13170, 13171, 13172, 13173,

13100, 13157, 13158, 13159, 13160, 13161, 13162, 13163, 13164, 13344, 13103, 13216, 13217,

13218, 13219, 13220, 13221, 13222, 13223, 13224, 13307, 13308, 13309, 13310, 13345, 13346,

13106, 13267, 13268, 13269, 13270, 13271, 13272, 13273, 13998, 13974, 13980, and 13274., or at least 95% identical to any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166, 13167,

13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158, 13159, 13160, 13161, 13162,

13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220, 13221, 13222, 13223, 13224,

13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268, 13269, 13270, 13271, 13272,

13273, 13998, 13974, 13980, and 13274. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099, 13165, 13166,

13272, 13273, 13998, 13974, 13980, and 13274., or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341, 13099,

13165, 13166, 13167, 13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158, 13159,

13160, 13161, 13162, 13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220, 13221,

13222, 13223, 13224, 13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268, 13269,

13270, 13271, 13272, 13273, 13998, 13974, 13980, and 13274., or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13329, 13334, 13335, 13091, 13259, 13260, 13261, 13262, 13263, 13264, 13265, 13266, 13336, 13311, 13312, 13341,

13099, 13165, 13166, 13167, 13168, 13169, 13170, 13171, 13172, 13173, 13100, 13157, 13158,

13159, 13160, 13161, 13162, 13163, 13164, 13344, 13103, 13216, 13217, 13218, 13219, 13220,

13221, 13222, 13223, 13224, 13307, 13308, 13309, 13310, 13345, 13346, 13106, 13267, 13268,

13269, 13270, 13271, 13272, 13273, 13998, 13974, 13980, and 13274. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. The sense strand may comprise an overhang. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence. In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662,

13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492, 13493, 13494, 13421, 13478, 13479,

13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424, 13537, 13538, 13539, 13540, 13541,

13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631, 13666, 13667, 13427, 13588, 13589,

13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or 13595. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492, 13493,

13494, 13421, 13478, 13479, 13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424, 13537,

13538, 13539, 13540, 13541, 13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631, 13666,

13667, 13427, 13588, 13589, 13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or 13595, at least 80% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488,

13489, 13490, 13491, 13492, 13493, 13494, 13421, 13478, 13479, 13480, 13481, 13482, 13483,

13484, 13485, 13665, 13424, 13537, 13538, 13539, 13540, 13541, 13542, 13543, 13544, 13545,

13628, 13629, 13630, 13631, 13666, 13667, 13427, 13588, 13589, 13590, 13591, 13592, 13593,

13594, 14126, 14102, 14108, or 13595, at least 85% identical to of any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632,

13633, 13662, 13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492, 13493, 13494, 13421,

13478, 13479, 13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424, 13537, 13538, 13539,

13540, 13541, 13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631, 13666, 13667, 13427,

13588, 13589, 13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or 13595, at least 90% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488, 13489,

13490, 13491, 13492, 13493, 13494, 13421, 13478, 13479, 13480, 13481, 13482, 13483, 13484,

13485, 13665, 13424, 13537, 13538, 13539, 13540, 13541, 13542, 13543, 13544, 13545, 13628,

13629, 13630, 13631, 13666, 13667, 13427, 13588, 13589, 13590, 13591, 13592, 13593, 13594,

14126, 14102, 14108, or 13595, or at least 95% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633,

13662, 13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492, 13493, 13494, 13421, 13478,

13479, 13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424, 13537, 13538, 13539, 13540,

13541, 13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631, 13666, 13667, 13427, 13588,

13589, 13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or 13595. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492, 13493, 13494,

13421, 13478, 13479, 13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424, 13537, 13538,

13539, 13540, 13541, 13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631, 13666, 13667,

13427, 13588, 13589, 13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or 13595, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488, 13489, 13490, 13491, 13492,

13493, 13494, 13421, 13478, 13479, 13480, 13481, 13482, 13483, 13484, 13485, 13665, 13424,

13537, 13538, 13539, 13540, 13541, 13542, 13543, 13544, 13545, 13628, 13629, 13630, 13631,

13666, 13667, 13427, 13588, 13589, 13590, 13591, 13592, 13593, 13594, 14126, 14102, 14108, or

13595, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13650, 13655, 13656, 13412, 13580, 13581, 13582, 13583, 13584, 13585, 13586, 13587, 13657, 13632, 13633, 13662, 13420, 13486, 13487, 13488, 13489, 13490,

13491, 13492, 13493, 13494, 13421, 13478, 13479, 13480, 13481, 13482, 13483, 13484, 13485,

13665, 13424, 13537, 13538, 13539, 13540, 13541, 13542, 13543, 13544, 13545, 13628, 13629,

13630, 13631, 13666, 13667, 13427, 13588, 13589, 13590, 13591, 13592, 13593, 13594, 14126,

14102, 14108, or 13595. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence.

[0073] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276, 13280, 13360,

13317, 13365, 13185, 13190, 13178, 13368, 13241, 13369, 13370, 13283, 13284, 13377, 13382,

13383, 13292, 13296, 13384, 13323, 13389, 13202, 13207, 13195, 13392, 13258, 13393, 13394,

13299, 14126, 14102, 14108, or 13300. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13329, 13334, 13335,

13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267,

13268, 13353, 13358, 13359, 13276, 13280, 13360, 13317, 13365, 13185, 13190, 13178, 13368,

13241, 13369, 13370, 13283, 13284, 13377, 13382, 13383, 13292, 13296, 13384, 13323, 13389,

13202, 13207, 13195, 13392, 13258, 13393, 13394, 13299, 14126, 14102, 14108, or 13300, at least 80% identical to any one of SEQ ID NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276,

13280, 13360, 13317, 13365, 13185, 13190, 13178, 13368, 13241, 13369, 13370, 13283, 13284,

13377, 13382, 13383, 13292, 13296, 13384, 13323, 13389, 13202, 13207, 13195, 13392, 13258,

13393, 13394, 13299, 14126, 14102, 14108, or 13300, at least 85% identical to of any one of SEQ ID

NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276, 13280, 13360, 13317, 13365,

13185, 13190, 13178, 13368, 13241, 13369, 13370, 13283, 13284, 13377, 13382, 13383, 13292,

13296, 13384, 13323, 13389, 13202, 13207, 13195, 13392, 13258, 13393, 13394, 13299, 14126,

14102, 14108, or 13300, at least 90% identical to any one of SEQ ID NOs 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267,

13202, 13207, 13195, 13392, 13258, 13393, 13394, 13299, 14126, 14102, 14108, or 13300, or at least 95% identical to any one of SEQ ID NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276,

13393, 13394, 13299, 14126, 14102, 14108, or 13300. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13329, 13334, 13335,

13202, 13207, 13195, 13392, 13258, 13393, 13394, 13299, 14126, 14102, 14108, or 13300, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276, 13280, 13360, 13317, 13365,

14102, 14108, or 13300, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13329, 13334, 13335, 13260, 13264, 13336, 13311, 13341, 13168, 13173, 13161, 13344, 13224, 13345, 13346, 13267, 13268, 13353, 13358, 13359, 13276,

13393, 13394, 13299, 14126, 14102, 14108, or 13300. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. The sense strand may comprise an overhang. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand.

[0074] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588, 13589, 13674, 13679, 13680, 13597,

13601, 13681, 13638, 13686, 13506, 13511, 13499, 13689, 13562, 13690, 13691, 13604, 13605,

13698, 13703, 13704, 13613, 13617, 13705, 13644, 13710, 13523, 13528, 13516, 13713, 13579,

13714, 13715, 13620, 14126, 14102, 14108, or 13621. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13650,

13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666,

13667, 13588, 13589, 13674, 13679, 13680, 13597, 13601, 13681, 13638, 13686, 13506, 13511,

13499, 13689, 13562, 13690, 13691, 13604, 13605, 13698, 13703, 13704, 13613, 13617, 13705,

13644, 13710, 13523, 13528, 13516, 13713, 13579, 13714, 13715, 13620, 14126, 14102, 14108, or 13621, at least 80% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588, 13589, 13674, 13679,

13680, 13597, 13601, 13681, 13638, 13686, 13506, 13511, 13499, 13689, 13562, 13690, 13691,

13604, 13605, 13698, 13703, 13704, 13613, 13617, 13705, 13644, 13710, 13523, 13528, 13516,

13713, 13579, 13714, 13715, 13620, 14126, 14102, 14108, or 13621, at least 85% identical to of any one of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588, 13589, 13674, 13679, 13680, 13597, 13601, 13681,

13638, 13686, 13506, 13511, 13499, 13689, 13562, 13690, 13691, 13604, 13605, 13698, 13703,

13704, 13613, 13617, 13705, 13644, 13710, 13523, 13528, 13516, 13713, 13579, 13714, 13715,

13620, 14126, 14102, 14108, or 13621, at least 90% identical to any one of SEQ ID NOs: 13650,

13644, 13710, 13523, 13528, 13516, 13713, 13579, 13714, 13715, 13620, 14126, 14102, 14108, or 13621, or at least 95% identical to any one of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585,

13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588, 13589, 13674,

13679, 13680, 13597, 13601, 13681, 13638, 13686, 13506, 13511, 13499, 13689, 13562, 13690,

13691, 13604, 13605, 13698, 13703, 13704, 13613, 13617, 13705, 13644, 13710, 13523, 13528,

13516, 13713, 13579, 13714, 13715, 13620, 14126, 14102, 14108, or 13621. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545,

13666, 13667, 13588, 13589, 13674, 13679, 13680, 13597, 13601, 13681, 13638, 13686, 13506,

13511, 13499, 13689, 13562, 13690, 13691, 13604, 13605, 13698, 13703, 13704, 13613, 13617,

13705, 13644, 13710, 13523, 13528, 13516, 13713, 13579, 13714, 13715, 13620, 14126, 14102,

14108, or 13621, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588, 13589, 13674, 13679, 13680, 13597,

13714, 13715, 13620, 14126, 14102, 14108, or 13621, or an antisense strand sequence thereof having

1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13650, 13655, 13656, 13581, 13585, 13657, 13632, 13662, 13489, 13494, 13482, 13665, 13545, 13666, 13667, 13588,

13589, 13674, 13679, 13680, 13597, 13601, 13681, 13638, 13686, 13506, 13511, 13499, 13689,

13562, 13690, 13691, 13604, 13605, 13698, 13703, 13704, 13613, 13617, 13705, 13644, 13710,

13523, 13528, 13516, 13713, 13579, 13714, 13715, 13620, 14126, 14102, 14108, or 13621. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence.

[0075] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 13998. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 13998, at least 80% identical to any one of SEQ ID NO: 13998, at least 85% identical to of any one of SEQ ID NO: 13998, at least 90% identical to any one of SEQ ID No: 13998, or at least 95% identical to any one of SEQ ID NO: 13998. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13998, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13998, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 13998. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 13998. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise an LDL receptor ligand. [0076] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NO: 14126. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 14126, at least 80% identical to any one of SEQ ID NO: 14126, at least 85% identical to of any one of SEQ ID NO: 14126, at least 90% identical to any one of SEQ ID NO: 14126, or at least 95% identical to any one of SEQ ID NO: 14126. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14126, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14126, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 14126. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand.

[0077] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 13974. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 13974, at least 80% identical to any one of SEQ ID NO: 13974, at least 85% identical to of any one of SEQ ID NO: 13974, at least 90% identical to any one of SEQ ID No: 13998, or at least 95% identical to any one of SEQ ID NO: 13974. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13974, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13974, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 13974. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 13974. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise an LDL receptor ligand. [0078] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NO: 14102. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 14102, at least 80% identical to any one of SEQ ID NO: 14102, at least 85% identical to of any one of SEQ ID NO: 14102, at least 90% identical to any one of SEQ ID NO: 14102, or at least 95% identical to any one of SEQ ID NO: 14102. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14102, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14102, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 14102. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand.

[0079] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 13980. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 13980, at least 80% identical to any one of SEQ ID NO: 13980, at least 85% identical to of any one of SEQ ID NO: 13980, at least 90% identical to any one of SEQ ID No: 13980, or at least 95% identical to any one of SEQ ID NO: 13980. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13980, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 13980, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 13980. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise a modification pattern described herein. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 13980. The sense strand may comprise an overhang. In some embodiments, the sense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The sense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise an LDL receptor ligand. [0080] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NO: 14108. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NO: 14108, at least 80% identical to any one of SEQ ID NO: 14108, at least 85% identical to of any one of SEQ ID NO: 14126, at least 90% identical to any one of SEQ ID NO: 14108, or at least 95% identical to any one of SEQ ID NO: 14108. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14108, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NO: 14108, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 14108. In some embodiments, the antisense strand further comprises a 3’ overhang. In some embodiments, the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the antisense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety, a GalNAc moiety, an integrin or an integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. The antisense strand may comprise a lipid moiety or a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise an LDL receptor ligand.

B. ASOs

[0081] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an antisense oligonucleotide (ASO). In some embodiments, the ASO is 12-30 nucleosides in length. In some embodiments, the ASO is 14-30 nucleosides in length. In some embodiments, the ASO is at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers. In some embodiments, the ASO is 15-25 nucleosides in length. In some embodiments, the ASO is 20 nucleosides in length.

[0082] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an ASO about 12-30 nucleosides in length and comprising a nucleoside sequence complementary to about 12-30 contiguous nucleosides of a full-length human GPAM mRNA sequence such as SEQ ID NO: 12867; wherein (i) the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified internucleoside linkage, and/or (ii) the composition comprises a pharmaceutically acceptable carrier. In some embodiments, the ASO comprise a nucleoside sequence complementary to at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more contiguous nucleosides of one of SEQ ID NO: 12867.

C. Modification patterns

[0083] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified internucleoside linkage, and/or (ii) the composition comprises a pharmaceutically acceptable carrier. In some embodiments, the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified internucleoside linkage. In some embodiments, the oligonucleotide comprises a modified intemucleoside linkage. In some embodiments, the modified internucleoside linkage comprises alkylphosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate tri ester, acetamidate, or carboxymethyl ester, or a combination thereof. In some embodiments, the modified internucleoside linkage comprises one or more phosphorothioate linkages. A phosphorothioate may include a nonbridging oxygen atom in a phosphate backbone of the oligonucleotide that is replaced by sulfur. Modified internucleoside linkages may be included in siRNAs or AS Os. Benefits of the modified intemucleoside linkage may include decreased toxicity or improved pharmacokinetics.

[0084] In some embodiments, the oligonucleotide comprises a duplex consisting of 21-36 nucleotide single strands with base pairing between 17-25 of the base pairs. In some embodiments, the duplex comprises blunt-ends at the 5 ’or 3’ ends of each strand. One strand (antisense strand) is complementary to a target mRNA. Each end of the antisense strand has one to five phosphorothioate bonds. The 5’ end has an optional phosphate mimic such as a vinyl phosphonate. In some embodiments, the oligonucleotide is used to knock down a target mRNA or a target protein. In some embodiments, the sense strand has the same sequence as the target mRNA. In some embodiments, there are 1-5 phosphorothi oates at the 5’ and 3’ ends.

[0085] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a modified intemucleoside linkage, wherein the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modified internucleoside linkages, or a range of modified internucleoside linkages defined by any two of the aforementioned numbers. In some embodiments, the oligonucleotide comprises no more than 18 modified intemucleoside linkages. In some embodiments, the oligonucleotide comprises no more than 20 modified intemucleoside linkages. In some embodiments, the oligonucleotide comprises 2 or more modified intemucleoside linkages, 3 or more modified intemucleoside linkages, 4 or more modified intemucleoside linkages, 5 or more modified intemucleoside linkages, 6 or more modified intemucleoside linkages, 7 or more modified intemucleoside linkages, 8 or more modified intemucleoside linkages, 9 or more modified intemucleoside linkages, 10 or more modified intemucleoside linkages, 11 or more modified intemucleoside linkages, 12 or more modified intemucleoside linkages, 13 or more modified intemucleoside linkages, 14 or more modified intemucleoside linkages, 15 or more modified intemucleoside linkages, 16 or more modified intemucleoside linkages, 17 or more modified intemucleoside linkages, 18 or more modified intemucleoside linkages, 19 or more modified intemucleoside linkages, or 20 or more modified intemucleoside linkages.

[0086] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises the modified nucleoside. In some embodiments, the modified nucleoside comprises a locked nucleic acid (LNA), hexitol nucleic acid (HNA), cyclohexene nucleic acid (CeNA), 2'-O-methoxy ethyl, 2'-O-alkyl, 2'-O-allyl, 2’-C-allyl, 2'- fluoro, or 2'-deoxy, or a combination thereof. In some embodiments, the modified nucleoside comprises a LNA. In some embodiments, the modified nucleoside comprises a 2’, 4’ constrained ethyl nucleic acid. In some embodiments, the modified nucleoside comprises HLA. In some embodiments, the modified nucleoside comprises CeNA. In some embodiments, the modified nucleoside comprises a 2'-methoxyethyl group. In some embodiments, the modified nucleoside comprises a 2'-O- methoxy ethyl group (“MOE”). In some embodiments, the modified nucleoside comprises a 2'-C-alkyl group. In some embodiments, the modified nucleoside comprises 2’ -methoxyethyl. For example, position 4 of the sense strand may comprise a methoxyethyl nucleoside such as a 2’ -O-methoxyethyl thymine. In some embodiments, the modified nucleoside comprises 2'-O-methyl. In some embodiments, the modified nucleoside comprises a 2'-O-allyl group. In some embodiments, the modified nucleoside comprises a 2'-fluoro group. In some embodiments, the modified nucleoside comprises a 2'-deoxy group. In some embodiments, the modified nucleoside comprises a 2'-O-methyl nucleoside, 2'- deoxy fluoro nucleoside, 2'-O-N-methylacetamido (2'-O-NMA) nucleoside, a 2'-O- dimethylaminoethoxyethyl (2'-O-DMAEOE) nucleoside, 2'-O-aminopropyl (2'-O-AP) nucleoside, or 2'-ara-F, or a combination thereof. In some embodiments, the modified nucleoside comprises a 2'-O- methyl nucleoside. In some embodiments, the modified nucleoside comprises a 2'-deoxyfluoro nucleoside. In some embodiments, the modified nucleoside comprises a 2'-O-NMA nucleoside. In some embodiments, the modified nucleoside comprises a 2'-O-DMAEOE nucleoside. In some embodiments, the modified nucleoside comprises a 2'-O-aminopropyl (2-O-AP) nucleoside. In some embodiments, the modified nucleoside comprises 2'-ara-F. In some embodiments, the modified nucleoside comprises one or more 2’ -fluoro modified nucleosides. In some embodiments, the modified nucleoside comprises a 2'-O-alkyl modified nucleoside. In some embodiments, the modified nucleoside comprises a 2’-O-methyl inosine nucleoside. In some embodiments, the modified nucleoside comprises an acyclic nucleic acid. In some embodiments, the acyclic nucleic is a glycol nucleic acid. In some embodiments, the modified nucleoside comprises an unlocked nucleic acid. Benefits of the modified nucleoside may include decreased toxicity or improved pharmacokinetics. [0087] In some embodiments, the modified nucleoside comprises a glycol nucleic acid (GNA). A GNA may comprise the following structure:

[0088] In some embodiments, the modified nucleoside comprises an unlocked nucleic acid. An unlocked nucleic acid may comprise the following structure:

wherein the base can be any pyrimidine or purine.

[0089] In some embodiments, the oligonucleotide comprises a modified nucleoside. In some embodiments, the modified nucleoside comprises a locked nucleic acid and an abasic site:

where J and K are independently an H or a 3’ or 5’ linkage to a nucleotide via a phosphodi ester or phosphorothioate bond.

[0090] In some embodiments, the oligonucleotide comprises a phosphate mimic. In some embodiments, the phosphate mimic comprises methylphosphonate. An example of a nucleotide that comprises a methylphosphonate is shown below:

(5’ methylphosphonate 2’-O-methyl Uridine).

[0091] In some embodiments, the oligonucleotide comprises a duplex consisting of 21 -36 nucleotide single strands with base pairing between 17-25 of the base pairs. In some embodiments, the duplex comprises blunt-ends at the 5 ’or 3’ ends of each strand. One strand (antisense strand) is complementary to a target mRNA. Each end of the antisense strand has one to five phosphorothioate bonds. The 5’ end has an optional phosphate mimic such as a vinyl phosphonate. In some embodiments, the oligonucleotide is used to knock down a target mRNA or a target protein. In some embodiments, the sense strand has the same sequence as the target mRNA. In some embodiments, there are 1-5 phosphorothi oates at the 5’ and 3’ ends.

[0092] In some embodiments, the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 modified nucleosides, or a range of nucleosides defined by any two of the aforementioned numbers. In some embodiments, the oligonucleotide comprises no more than 19 modified nucleosides. In some embodiments, the oligonucleotide comprises no more than 21 modified nucleosides. In some embodiments, the oligonucleotide comprises 2 or more modified nucleosides, 3 or more modified nucleosides, 4 or more modified nucleosides, 5 or more modified nucleosides, 6 or more modified nucleosides, 7 or more modified nucleosides, 8 or more modified nucleosides, 9 or more modified nucleosides, 10 or more modified nucleosides, 11 or more modified nucleosides, 12 or more modified nucleosides, 13 or more modified nucleosides, 14 or more modified nucleosides, 15 or more modified nucleosides, 16 or more modified nucleosides, 17 or more modified nucleosides, 18 or more modified nucleosides, 19 or more modified nucleosides, 20 or more modified nucleosides, or 21 or more modified nucleosides.

[0093] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a moiety attached at a 3’ or 5’ terminus of the oligonucleotide. Examples of moieties include a hydrophobic moiety or a sugar moiety, or a combination thereof. In some embodiments, the oligonucleotide is an siRNA having a sense strand, and the moiety is attached to a 5’ end of the sense strand. In some embodiments, the oligonucleotide is an siRNA having a sense strand, and the moiety is attached to a 3’ end of the sense strand. In some embodiments, the oligonucleotide is an siRNA having an antisense strand, and the moiety is attached to a 5’ end of the antisense strand. In some embodiments, the oligonucleotide is an siRNA having an antisense strand, and the moiety is attached to a 3’ end of the antisense strand. In some embodiments, the oligonucleotide is an ASO, and the moiety is attached to a 5’ end of the ASO. In some embodiments, the oligonucleotide is an ASO, and the moiety is attached to a 3’ end of the ASO. [0094] In some embodiments, the sense strand comprises at least three modified nucleosides, wherein the three modifications comprise a 2’ -fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O-methoxyethyl. In some embodiments, the sense strand comprises at least two modified nucleosides, wherein the two modifications comprise a 2’ -fluoro modified nucleoside, a 2’- O-methyl modified nucleoside, and 2’-O-methoxyethyl. In some embodiments, each nucleoside of the sense strand comprises a modified nucleoside, wherein the modified nucleosides are selected from the group consisting of a 2’ -fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O- methoxy ethyl. In some embodiments, the sense strand comprises at least a 2’ -fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O-methoxyethyl.

[0095] In some embodiments, the antisense strand is combination of 2’ -fluoro and 2’-O-methyl modifications. In some embodiments, each nucleoside of the antisense strand comprises a modified nucleoside, wherein the modified nucleosides are selected from the group consisting of a 2’ -fluoro modified nucleoside and a2’-O-methyl modified nucleoside. In some embodiments, the sense strand comprises at least a 2’ -fluoro modified nucleoside and a 2’-O-methyl modified nucleoside.

[0096] The oligonucleotide may include purines. Examples of purines include adenine (A), inosine (I), or guanine (G), or modified versions thereof. The oligonucleotide may include pyrimidines. Examples of pyrimidines include cytosine (C), thymine (T), or uracil (U), or modified versions thereof.

[0097] In some embodiments, the sense strand comprises purines and pyrimidines. In some embodiments, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’ -O-methyl and 2’ -O-methoxyethyl. In some embodiments, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O-methoxyethyl. In some embodiments, all purine nucleosides comprise 2’ -O-methoxyethyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl. In some embodiments, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -O-methyl and 2’ -O-methoxyethyl. In some embodiments, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methoxyethyl. In some embodiments, all pyrimidine nucleosides comprise 2’-O- methoxy ethyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl. In some embodiments, the sense strand may include a 2’ -deoxy nucleoside.

[0098] In some embodiments, at least one nucleotide at position 4 or 5 of the sense strand comprises a 2’ -O-methoxyethyl modified nucleoside. In some embodiments, at least one nucleotide of the sense strand from position 6 to 9 comprise a 2’-fluoro-modified nucleoside. In some embodiments, at least two nucleotides of the sense strand at position 6 to 9 comprise a 2’-fluoro-modified nucleoside. In some embodiments, at least three nucleotides of the sense strand at positions 6 to 9 comprise a 2’ - fluoro-modified nucleoside. In some embodiments, each nucleotide from positions 6 to 9 of the sense strand comprise a 2’ -fluoro-modified nucleoside. In some embodiments, at least one nucleotide at position 16 to 20 of the sense strand comprises a 2’-O-methyl modified nucleoside. In some embodiments, at least two nucleotides at position 16 to 20 of the sense strand comprise a 2’ -O-methyl modified nucleoside. In some embodiments, at least three nucleotides at position 16 to 20 of the sense strand comprise a 2’ -O-methyl modified nucleoside. In some embodiments, at least four nucleotides at position 16 to 20 of the sense strand comprise a 2’ -O-methyl modified nucleoside. In some embodiments, all nucleotides at position 16 to 20 of the sense strand comprise a 2’ -O-methyl modified nucleoside.

[0099] In some embodiments, any of the following is true with regards to the antisense strand: all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’ -O-methyl; all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl; all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides comprise 2’ -fluoro; all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O- methyl; all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl; or all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides comprise 2’ -fluoro. In some embodiments, all purine nucleosides comprise 2’ - fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl. In some embodiments, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl; all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides comprise 2’ -fluoro. In some embodiments, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O- methyl; all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl. In some embodiments, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides comprise 2’ -fluoro.

[0100] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a hydrophobic moiety. The hydrophobic moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide. The hydrophobic moiety may include a lipid such as a fatty acid. The hydrophobic moiety may include a hydrocarbon. The hydrocarbon may be linear. The hydrocarbon may be non-linear. The hydrophobic moiety may include a lipid moiety or a cholesterol moiety, or a combination thereof.

[0101] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a lipid attached at a 3’ or 5’ terminus of the oligonucleotide. In some embodiments, the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl stearyl, or a-tocopherol, or a combination thereof.

[0102] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a sugar moiety. The sugar moiety may include an N- acetyl galactose moiety (e.g., anN-acetylgalactosamine (GalNAc) moiety), an N-acetyl glucose moiety (e.g., an N-acetylglucosamine (GlcNAc) moiety), a fucose moiety, or a mannose moiety. The sugar moiety may include 1, 2, 3, or more sugar molecules. The sugar moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide. The sugar moiety may include an N-acetyl galactose moiety. The sugar moiety may include an N-acetylgalactosamine (GalNAc) moiety. The sugar moiety may include an N-acetyl glucose moiety. The sugar moiety may include N- acetylglucosamine (GlcNAc) moiety. The sugar moiety may include a fucose moiety. The sugar moiety may include a mannose moiety. N-acetyl glucose, GlcNAc, fucose, or mannose may be useful for targeting macrophages since they may target or bind a mannose receptor such as CD206.

[0103] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) moiety. GalNAc may be useful for hepatocyte targeting. The GalNAc moiety may include a bivalent or trivalent branched linker. The oligo may be attached to 1, 2 or 3 GalNAcs through a bivalent or trivalent branched linker. The GalNAc moiety may include 1, 2, 3, or more GalNAc molecules. The GalNAc moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.

[0104] The oligonucleotide may include purines. Examples of purines include adenine (A), inosine (I), guanine (G), or modified versions thereof. The oligonucleotide may include pyrimidines. Examples of pyrimidines include cytosine (C), thymine (T), or uracil (U), or modified versions thereof.

[0105] In some embodiments, purines of the oligonucleotide comprise 2’ -fluoro modified purines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines. In some embodiments, all purines of the oligonucleotide comprise 2’ -fluoro modified purines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, all purines of the oligonucleotide comprise a mixture of 2’ - fluoro and 2’-O-methyl modified purines. 2’-O-methyl may include 2’-O-methyl. Where 2’-O-methyl modifications are described, it is contemplated that a 2’ -methyl modification may be included, and vice versa.

[0106] In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. [0107] In some embodiments, purines of the oligonucleotide comprise 2’-fluoro modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’ -O-methyl modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’ -fluoro modified purines, and pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’ -O-methyl modified purines, and pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and purines of the oligonucleotide comprise 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines, and purines of the oligonucleotide comprise 2’ -fluoro modified purines.

[0108] In some embodiments, all purines of the oligonucleotide comprise 2’ -fluoro modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’ -O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’ -fluoro modified purines, and all pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’ -O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and all purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines, and all purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and all purines of the oligonucleotide comprise 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -O-methyl modified pyrimidines, and all purines of the oligonucleotide comprise 2’ -fluoro modified purines.

[0109] In some embodiments, purines of the oligonucleotide comprise 2’ -fluoro modified purines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all purines of the oligonucleotide comprise 2 ’-fluoro modified purines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, all purines of the oligonucleotide comprise a mixture of 2’- fluoro and 2’-O-methyl modified purines. 2’-O-methyl may include 2’-O-methyl.

[0110] In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines.

[0111] In some embodiments, purines of the oligonucleotide comprise 2’-fluoro modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’ -fluoro modified purines, and pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines, and pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-fluoro modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and purines of the oligonucleotide comprise 2’ -fluoro modified purines.

[0112] In some embodiments, all purines of the oligonucleotide comprise 2’-fluoro modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’ -fluoro modified purines, and all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise 2’-fluoro modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and all purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimi dines, and all purines of the oligonucleotide comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and all purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and all purines of the oligonucleotide comprise 2’ -fluoro modified purines.

[0113] In some cases, the oligonucleotide comprises a particular modification pattern. In some embodiments, position 9 counting from the 5’ end of the of a strand of the oligonucleotide may have a 2’F modification. In some embodiments, when position 9 of a strand of the oligonucleotide is a pyrimidine, then all purines in a strand of the oligonucleotide have a 2’OMe modification. In some embodiments, when position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in a strand of the oligonucleotide. In some embodiments, when position 9 and only one other base between positions 5 and 11 of a strand of the oligonucleotide are pyrimidines, then both of these pyrimidines are the only two positions with a 2’F modification in a strand of the oligonucleotide. In some embodiments, when position 9 and only two other bases between positions 5 and 11 of a strand of the oligonucleotide are pyrimidines, and those two other pyrimidines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. In some embodiments, when there are more than 2 pyrimidines between positions 5 and 11 of a strand of the oligonucleotide, then all combinations of pyrimidines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that a strand of the oligonucleotide does not have three 2’F modifications in a row. In some cases, a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to any or all of these a strand of the oligonucleotide rules.

[0114] In some embodiments, when position 9 of a strand of the oligonucleotide is a purine, then all purines in a strand of the oligonucleotide have a 2’OMe modification. In some embodiments, when position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in a strand of the oligonucleotide. In some embodiments, when position 9 and only one other base between positions 5 and 11 of a strand of the oligonucleotide are purines, then both of these purines are the only two positions with a 2’F modification in a strand of the oligonucleotide. In some embodiments, when position 9 and only two other bases between positions 5 and 11 of a strand of the oligonucleotide are purines, and those two other purines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. In some embodiments, when there are more than 2 purines between positions 5 and 11 of a strand of the oligonucleotide, then all combinations of purines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that a strand of the oligonucleotide does not have three 2’F modifi cations in a row. In some cases, a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to any or all of these a strand of the oligonucleotide rules. [0115] In some cases, position 9 of a strand of the oligonucleotide can be a 2’ deoxy. In these cases, 2’F and 2’OMe modifications may occur at the other positions of a strand of the oligonucleotide. In some cases, a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to these a strand of the oligonucleotide rules.

[0116] In some embodiments, the oligonucleotide is delivered to a cell or tissue by linking the oligonucleotide to a targeting group. In some embodiments, the targeting group includes a cell receptor ligand, such as an integrin targeting ligand. Integrins may include a family of transmembrane receptors that facilitate cell-extracellular matrix (ECM) adhesion. In some embodiments, the moiety includes an epithelial-specific integrin. Integrin alpha-v-beta-6 (αvβ6) bay be an example of an epithelial-specific integrin αvβ6 may be a receptor for an ECM protein or TGF-beta latency- associated peptide (LAP). Integrin αvβ6 may be expressed in a cell or tissue. Integrin αvβ6 may be expressed or upregulated in injured pulmonary epithelium.

[0117] In some embodiments, the oligonucleotide is linked to an integrin targeting ligand that has affinity for integrin αvβ6. An integrin targeting ligand may include a compound that has affinity for integrin αvβ6 or integrin alpha- v-beta-3 (αvβ3), may be useful as a ligand to facilitate targeting or delivery of the oligonucleotide to which it is attached to a particular cell type or tissue (e.g., to cells expressing integrin αvβ3 or αvβ6). In some embodiments, multiple integrin targeting ligands are linked to the oligonucleotide. In some embodiments, the oligonucleotide-integrin targeting ligand conjugates are selectively internalized by chondrocytes, either through receptor-mediated endocytosis or by other means.

[0118] In some embodiments, an oligonucleotide that targets GPAM further comprises a targeting ligand that targets a receptor which mediates delivery to a specific CNS tissue. In some embodiments, the targeting ligand is conjugated to the oligonucleotide. In one embodiment, the targeting ligand is selected from the group consisting of Angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand. In one embodiment, the targeting ligand is selected from the group consisting of trans-retinol, RGD peptide, LDL receptor ligand, and carbohydrate-based ligands. In one embodiment, the targeting ligand is a RGD peptide, such as H-Gly-Arg-Gly-Asp-Ser- Pro-Lys-Cys-OH or Cyclo(-Arg-Gly-Asp-D-Phe-Cys).

[0119] Examples of targeting groups useful for delivering the oligonucleotide that include integrin targeting ligands may be based upon peptides or peptide mimics containing an arginine-glycine- aspartic acid (RGD) peptide. In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an RGD peptide. In some embodiments, the composition comprises an RGD peptide. In some embodiments, the composition comprises an RGD peptide derivative. In some embodiments, the RGD peptide is attached at a 3’ terminus of the oligonucleotide. In some embodiments, the RGD peptide is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the composition comprises a sense strand, and the RGD peptide is attached to the sense strand (e.g., attached to a 5’ end of the sense strand, or attached to a 3’ end of the sense strand). In some embodiments, the composition comprises an antisense strand, and the RGD peptide is attached to the antisense strand (e.g., attached to a 5’ end of the antisense strand, or attached to a 3’ end of the antisense strand). In some embodiments, the composition comprises an RGD peptide attached at a 3’ or 5’ terminus of the oligonucleotide. In some embodiments, the oligonucleotide comprises an RGD peptide, and a lipid attached at a 3’ or 5’ terminus of the oligonucleotide. The RGD peptide may be linear. The RGD peptide may be cyclic. An RGD peptide may include a D-amino acid. In some embodiments, the RGD peptide comprises Cyclo(-Arg-Gly-Asp-D-Phe-Cys). In some embodiments, the RGD peptide comprises Cyclo(-Arg- Gly-Asp-D-Phe-Lys). In some embodiments, the RGD peptide comprises Cyclo(-Arg-Gly-Asp-D- Phe-azido). In some embodiments, the RGD peptide comprises an amino benzoic acid derived RGD. In some embodiments, the RGD peptide comprises Cyclo(-Arg-Gly-Asp-D-Phe-Cys), Cyclo(-Arg- Gly-Asp-D-Phe-Lys), Cyclo(-Arg-Gly-Asp-D-Phe-azido), an amino benzoic acid derived RGD, or a combination thereof. In some embodiments, the RGD peptide comprises multiple of such RGD peptides. For example, the RGD peptide may include 2, 3, or 4 RGD peptides. Some embodiments include an arginine-glycine-glutamic acid peptide.

[0120] In some embodiments, position nine of the sense strand comprises a 2’-fluoro-modified pyrimidine. In some embodiments, all purines of the sense strand comprise 2’-O-methyl modified purines. In some embodiments, 1, 2, 3, 4, or 5 pyrimidines between positions 5 and 11 comprise a 2’- fluoro-modified pyrimidine, provided there are not three 2’ -fluoro- modified pyrimidines in a row. In some embodiments, the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotide. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides, 2’-O- methyl modified nucleotides and unmodified deoxyribonucleotide. In some embodiments, position nine of the sense strand comprises a 2’-fluoro-modified pyrimidine; all purines of the sense strand comprises 2’-O-methyl modified purines; 1, 2, 3, 4, or 5 pyrimidines between positions 5 and 11 comprise a 2’-fluoro-modified pyrimidine, provided there are not three 2’-fluoro-modified pyrimidines in a row; the odd-numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’-fluoro- modified nucleotides and unmodified deoxyribonucleotides.

[0121] In some embodiments, position nine of the sense strand comprises a 2’-fluoro-modified purine. In some embodiments, all pyrimidines of the sense strand comprise 2’ -O-methyl modified purines. In some embodiments, 1, 2, 3, 4, or 5 purines between positions 5 and 11 comprise a 2’- fluoro-modified purine, provided there are not three 2’-fluoro-modified purine in a row. In some embodiments, the odd-numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’- fluoro-modified nucleotides and unmodified deoxyribonucleotide. In some embodiments, the even- numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides, 2’ -O-methyl modified nucleotides and unmodified deoxyribonucleotide. In some embodiments, position nine of the sense strand comprises a 2’-fluoro-modified purine; all pyrimidine of the sense strand comprises 2’- O-methyl modified pyrimidines; 1, 2, 3, 4, or 5 purines between positions 5 and 11 comprise a 2’- fluoro-modified purines, provided there are not three 2’-fluoro-modified purines in a row; the odd- numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides; and the even- numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, there are not three 2’-fluoro-modified purines in a row. In some embodiments, there are not three 2’-fluoro-modified pyrimidines in a row.

[0122] In some embodiments, position nine of the sense strand comprises an unmodified deoxyribonucleotide. In some embodiments, positions 5, 7, and 8 of the sense strand comprise 2’- fluoro-modifed nucleotides. In some embodiments, all pyrimidines in positions 10 to 21 of the sense strand comprise 2’ -O-methyl modified pyrimidines and all purines in positions 10 to 21 of the comprise 2’ -O-methyl modified purines or 2’-fluoro-modified purines. In some embodiments, the odd-numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides, 2’ -O-methyl modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, position nine of the sense strand comprises an unmodified deoxyribonucleotide; positions 5, 7, and 8 of the sense strand comprise 2’-fluoro-modifed nucleotides; all pyrimidines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified pyrimidines and all purines in positions 10 to 21 of the comprise 2’- O-methyl modified purines or 2’-fluoro-modified purines; the odd-numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotides. [0123] In some embodiments, position nine of the sense strand comprises an unmodified deoxyribonucleotide. In some embodiments, positions 5, 7, and 8 of the sense strand comprise 2’- fluoro-modifed nucleotides. In some embodiments, all purines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified purines and all pyrimidines in positions 10 to 21 of the comprise 2’ - O-methyl modified pyrimidines or 2’-fluoro-modified pyrimidines. In some embodiments, the odd- numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides, 2’ -O-methyl modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, position nine of the sense strand comprises an unmodified deoxy ribonucleotide; positions 5, 7, and 8 of the sense strand comprise 2’-fluoro-modifed nucleotides; all purines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified purines and all pyrimidines in positions 10 to 21 of the comprise 2’ -O-methyl modified pyrimidines or 2’-fluoro-modified pyrimidines; the odd-numbered positions of the antisense strand comprise 2’ -O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’-fluoro-modified nucleotides and unmodified deoxyribonucleotide.

[0124] In some embodiments, the moiety includes a negatively charged group attached at a 5’ end of the oligonucleotide. This may be referred to as a 5 ’ -end group. In some embodiments, the negatively charged group is attached at a 5’ end of an antisense strand of an siRNA disclosed herein. The 5’ -end group may be or include a 5’ -end phosphorothioate, 5 ’-end phosphorodithioate, 5 ’-end vinylphosphonate (5 ’-VP), 5 ’-end methylphosphonate, 5 ’-end cyclopropyl phosphonate, or a 5’- deoxy-5’-C-malonyl. The 5’-end group may comprise 5’-VP. In some embodiments, the 5’-VP comprises a trans- vinylphosphonate or cis- vinylphosphonate. The 5 ’-end group may include an extra 5’ phosphate. A combination of 5 ’-end groups may be used.

[0125] In some embodiments, the oligonucleotide includes a negatively charged group. The negatively charged group may aid in cell or tissue penetration. The negatively charged group may be attached at a 5’ or 3’ end (e.g., a 5’ end) of the oligonucleotide. This may be referred to as an end group. The end group may be or include a phosphorothioate, phosphorodithioate, vinylphosphonate, methylphosphonate, cyclopropyl phosphonate, or a deoxy-C-malonyl. The end group may include an extra 5’ phosphate such as an extra 5’ phosphate. A combination of end groups may be used.

[0126] In some embodiments, the oligonucleotide includes a phosphate mimic. In some embodiments, the phosphate mimic comprises vinyl phosphonate. In some embodiments, the vinyl phosphonate comprises a trans- vinylphosphonate. In some embodiments, the vinyl phosphonate comprises a cis- vinylphosphonate. An example of a nucleotide that includes a vinyl phosphonate is shown below.

5’ vinylphosphonate 2’ -O-methyl Uridine

[0127] In some embodiments, the vinyl phosphonate increases the stability of the oligonucleotide. In some embodiments, the vinyl phosphonate increases the accumulation of the oligonucleotide in tissues. In some embodiments, the vinyl phosphonate protects the oligonucleotide from an exonuclease or a phosphatase. In some embodiments, the vinyl phosphonate improves the binding affinity of the oligonucleotide with the siRNA processing machinery.

[0128] In some embodiments, the oligonucleotide includes 1 vinyl phosphonate. In some embodiments, the oligonucleotide includes 2 vinyl phosphonates. In some embodiments, the oligonucleotide includes 3 vinyl phosphonates. In some embodiments, the oligonucleotide includes 4 vinyl phosphonates. In some embodiments, the antisense strand of the oligonucleotide comprises a vinyl phosphonate at the 5’ end. In some embodiments, the antisense strand of the oligonucleotide comprises a vinyl phosphonate at the 3’ end. In some embodiments, the sense strand of the oligonucleotide comprises a vinyl phosphonate at the 5’ end. In some embodiments, the sense strand of the oligonucleotide comprises a vinyl phosphonate at the 3’ end.

1. Hydrophobic moieties

[0129] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a hydrophobic moiety. The hydrophobic moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide. The hydrophobic moiety may include a lipid such as a fatty acid. The hydrophobic moiety may include a hydrocarbon. The hydrocarbon may be linear. The hydrocarbon may be non-linear. The hydrophobic moiety may include a lipid moiety or a cholesterol moiety, or a combination thereof.

[0130] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a lipid attached at a 3’ or 5’ terminus of the oligonucleotide. In some embodiments, the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl, stearyl, or a-tocopherol, or a combination thereof.

[0131] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a hydrophobic ligand or moiety. In some embodiments, the hydrophobic ligand or moiety comprises cholesterol. In some embodiments, the hydrophobic ligand or moiety comprises a cholesterol derivative. In some embodiments, the hydrophobic ligand or moiety is attached at a 3’ terminus of the oligonucleotide. In some embodiments, the hydrophobic ligand or moiety is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the composition comprises a sense strand, and the hydrophobic ligand or moiety is attached to the sense strand (e.g., attached to a 5’ end of the sense strand, or attached to a 3’ end of the sense strand). In some embodiments, the composition comprises an antisense strand, and the hydrophobic ligand or moiety is attached to the antisense strand (e.g., attached to a 5’ end of the antisense strand, or attached to a 3’ end of the antisense strand). In some embodiments, the composition comprises a hydrophobic ligand or moiety attached at a 3’ or 5’ terminus of the oligonucleotide. [0132] In some embodiments, ahydrophobic moiety is attached to the oligonucleotide (e.g., a sense strand and/or an antisense strand of a siRNA). In some embodiments, a hydrophobic moiety is attached at a 3’ terminus of the oligonucleotide. In some embodiments, a hydrophobic moiety is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the hydrophobic moiety comprises cholesterol. In some embodiments, the hydrophobic moiety includes a cyclohexanyl. The hydrophobic moiety may include an esterified lipid.

[0133] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a lipid attached at a 3’ or 5’ terminus of the oligonucleotide. In some embodiments, a lipid is attached at a 3’ terminus of the oligonucleotide. In some embodiments, a lipid is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl, stearyl, or a-tocopherol, or a combination thereof. In some embodiments, the lipid comprises stearyl, lithocholyl, docosanyl, docosahexaenyl, or myristyl. In some embodiments, the lipid comprises cholesterol. In some embodiments, the lipid includes a sterol such as cholesterol. In some embodiments, the lipid comprises stearyl, t-butylphenol, n-butylphenol, octylphenol, dodecylphenol, phenyl n-dodecyl, octadecylbenzamide, hexadecylbenzamide, or octadecylcyclohexyl. In some embodiments, the lipid comprises phenyl para C12. The hydrophobic moiety may include an esterified lipid.

[0134] In some embodiments, the oligonucleotide comprises any aspect of the following structure:

. In some embodiments, the oligonucleotide comprises any aspect of the following structure:

In some embodiments, the oligonucleotide comprises any aspect of the following structure:

In some embodiments, the oligonucleotide comprises any aspect of the following structure: The aspect included in the oligonucleotide may include the entire structure, or may include the lipid moiety, of any of the structures shown. In some embodiments, n is 1-3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, R is an alkyl group. In some embodiments, the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbons. In some embodiments, the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons. In some embodiments, the alkyl group contains 4-18 carbons. In some embodiments, the lipid moiety comprises an alcohol or ether.

[0135] In some embodiments, the lipid includes a fatty acid. In some embodiments, the lipid comprises a lipid depicted in Table 1. The example lipid moieties in Table 1 are shown attached at a 5’ end of an oligonucleotide, in which the 5’ terminal phosphate of the oligonucleotide is shown with the lipid moiety. In some embodiments, a lipid moiety in Table 1 may be attached at a different point of attachment than shown. For example, the point of attachment of any of the lipid moieties in the table may be at a 3’ oligonucleotide end. In some embodiments, the lipid is used for targeting the oligonucleotide to a non-hepatic cell or tissue.

Table 1: Hydrophobic moiety examples

[0136] In some embodiments, the lipid or lipid moiety includes 16 to 18 carbons. In some embodiments, the lipid includes 16 carbons. In some embodiments, the lipid includes 17 carbons. In some embodiments, the lipid includes 18 carbons. In some embodiments, the lipid moiety includes 16 carbons. In some embodiments, the lipid moiety includes 17 carbons. In some embodiments, the lipid moiety includes 18 carbons.

[0137] The hydrophobic moiety may include a linker that comprises a carbocycle. The carbocycle may be six-membered. Some examples of a carbocycle include phenyl or cyclohexyl. The linker may include a phenyl. The linker may include a cyclohexyl. The lipid may be attached to the carbocycle, which may in turn be attached at a phosphate (e. g. , 5 ’ or 3 ’ phosphate) of the oligonucleotide. In some embodiments, the lipid or hydrocarbon, and the end of the sense are connected to the phenyl or cyclohexyl linker in the 1,4; 1,3; or 1,2 substitution pattern (e.g., the para, meta, or ortho phenyl configuration). In some embodiments, the lipid or hydrocarbon, and the end of the sense are connected to the phenyl or cyclohexyl linker in the 1,4 substitution pattern (e.g., the para phenyl configuration). The lipid may be attached to the carbocycle in the 1,4 substitution pattern relative to the oligonucleotide. The lipid may be attached to the carbocycle in the 1,3 substitution pattern relative to the oligonucleotide. The lipid may be attached to the carbocycle in the 1,2 substitution pattern relative to the oligonucleotide. The lipid may be attached to the carbocycle in the ortho orientation relative to the oligonucleotide. The lipid may be attached to the carbocycle in the para orientation relative to the oligonucleotide. The lipid may be attached to the carbocycle in the meta orientation relative to the oligonucleotide.

[0138] The lipid moiety may comprise or consist of the following structure

. In some embodiments, the lipid moiety comprises or consists of the following structure:

In some embodiments, the lipid moiety comprises the following structure:

In some embodiments, the lipid moiety comprises or consist of the following structure:

. In some embodiments, the dotted line indicates a covalent connection. The covalent connection may between an end of the sense or antisense strand. For example, the connection may be to the 5’ end of the sense strand. In some embodiments, n is 0-3. In some embodiments, n is 1-3. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, Ris an alkyl group. In some embodiments, the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbons. In some embodiments, the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons. In some embodiments, R comprises or consists of an alkyl group containing 4-18 carbons.

[0139] The lipid moiety may be attached at a 5’ end of the oligonucleotide. The 5’ end may have one phosphate linking the lipid moiety to a 5’ carbon of a sugar of the oligonucleotide. The 5’ end may have two phosphates linking the lipid moiety to a 5’ carbon of a sugar of the oligonucleotide. The 5’ end may have three phosphates linking the lipid moiety to a 5’ carbon of a sugar of the oligonucleotide. The 5’ end may have one phosphate connected to the 5’ carbon of a sugar of the oligonucleotide, where the one phosphate is connected to the lipid moiety. The 5’ end may have two phosphates connected to the 5’ carbon of a sugar of the oligonucleotide, where the one of the two phosphates is connected to the lipid moiety. The 5’ end may have three phosphates connected to the 5’ carbon of a sugar of the oligonucleotide, where the one of the three phosphates is connected to the lipid moiety. The sugar may include a ribose. The sugar may include a deoxyribose. The sugar may be modified a such as a 2’ modified sugar (e.g., a 2’-O-methyl or 2’-fluoro ribose). A phosphate ofthe 5’ end may include a modification such as a sulfur in place of an oxygen. Two phosphates of the 5’ end may include a modification such as a sulfur in place of an oxygen. Three phosphates of the 5’ end may include a modification such as a sulfur in place of an oxygen.

[0140] In some embodiments, the oligonucleotide includes 1 lipid moiety. In some embodiments, the oligonucleotide includes 2 lipid moieties. In some embodiments, the oligonucleotide includes 3 lipid moieties. In some embodiments, the oligonucleotide includes 4 lipid moieties.

[0141] Some embodiments relate to a method of making an oligonucleotide comprising a hydrophobic conjugate. A strategy for making hydrophobic conjugates may include use of a phosphorami dite reagent based upon a 6-membered ring alcohol such as a phenol or cyclohexanol. The phosphoramidite may be reacted to a nucleotide to connect the nucleotide to the hydrophobic moiety, and thereby produce the hydrophobic conjugate. Some examples of phosphoramidite reagents that may be used to produce a hydrophobic conjugate are provided as follows:

or

In some embodiments, n is 1-3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, Ris an alkyl group. In some embodiments, the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or20 carbons. In some embodiments, the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons. In some embodiments, R comprises or consists of an alkyl group containing 4-18 carbons. Any one of the phosphorami dite reagents may be reacted to a 5’ end of an oligonucleotide to produce an oligonucleotide comprising a hydrophobic moiety. In some embodiments, the phosphorami dite reagents is reacted to a 5’ end of a sense strand of an siRNA. The sense strand may then be hybridized to an antisense strand to form a duplex. The hybridization may be performed by incubating the sense and antisense strands in solution at a given temperature. The temperature may be gradually reduced. The temperature may comprise or include a temperature comprising an annealing temperature for the sense and antisense strands. The temperature may be below or include a temperature below the annealing temperature for the sense and antisense strands. The temperature may be below a melting temperature of the sense and antisense strands.

[0142] The lipid may be attached to the oligonucleotide by a linker. The linker may include a polyethyleneglycol (e.g., tetraethyleneglycol).

[0143] The modifications described herein may be useful for delivery to a cell or tissue, for example, extrahepatic delivery or targeting of an oligonucleotide composition. The modifications described herein may be useful for targeting an oligonucleotide composition to a cell or tissue.

2. Sugar moieties

[0144] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a sugar moiety. The sugar moiety may include an N- acetyl galactose moiety (e.g., anN-acetylgalactosamine (GalNAc) moiety), an N-acetyl glucose moiety (e.g., an N-acetylglucosamine (GlcNAc) moiety), a fucose moiety, or a mannose moiety. The sugar moiety may include 1, 2, 3, or more sugar molecules. The sugar moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide. The sugar moiety may include an N-acetyl galactose moiety. The sugar moiety may include an N-acetylgalactosamine (GalNAc) moiety. The sugar moiety may include an N-acetyl glucose moiety. The sugar moiety may include N- acetylglucosamine (GlcNAc) moiety. The sugar moiety may include a fucose moiety. The sugar moiety may include a mannose moiety. N-acetyl glucose, GlcNAc, fucose, or mannose may be useful for targeting macrophages when they target or bind a mannose receptor such as CD206. The sugar moiety may be useful for binding or targeting an asialoglycoprotein receptor such as an asialoglycoprotein receptor of a hepatocyte. The GalNAc moiety may bind to an asialoglycoprotein receptor. The GalNAc moiety may target a hepatocyte.

[0145] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) moiety. GalNAc may be useful for hepatocyte targeting. The GalNAc moiety may include a bivalent or trivalent branched linker. The oligo may be attached to 1, 2 or 3 GalNAcs through a bivalent or trivalent branched linker. The GalNAc moiety may include 1, 2, 3, or more GalNAc molecules. The GalNAc moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.

[0146] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) ligand for hepatocyte targeting. In some embodiments, the composition comprises GalNAc. In some embodiments, the composition comprises a GalNAc derivative. In some embodiments, the GalNAc ligand is attached at a 3’ terminus of the oligonucleotide. In some embodiments, the GalNAc ligand is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the composition comprises a sense strand, and the GalNAc ligand is attached to the sense strand (e.g., attached to a 5’ end of the sense strand, or attached to a 3’ end of the sense strand). In some embodiments, the composition comprises an antisense strand, and the GalNAc ligand is attached to the antisense strand (e.g., attached to a 5’ end of the antisense strand, or attached to a 3’ end of the antisense strand). In some embodiments, the composition comprises a GalNAc ligand attached at a 3’ or 5’ terminus of the oligonucleotide.

[0147] Disclosed herein, in some embodiments, are compositions comprising an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises a GalNAc moiety. The GalNAc moiety may be included in any formula, structure, or GalNAc moiety shown below. In some embodiments, described herein is a compound (e.g., oligonucleotide) represented by Formula (I) or (II):

or a salt thereof, wherein

J is an oligonucleotide; each w is independently selected from any value from 1 to 20; each v is independently selected from any value from 1 to 20; n is selected from any value from 1 to 20; m is selected from any value from 1 to 20; z is selected from any value from 1 to 3, wherein if z is 3, Y is C if z is 2, Y is CR⁶, or if z is 1, Y is C(R⁶)₂;

Q is selected from:

C_3-10 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -NO₂, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂,-N(R⁷)C(O)R⁷ -N(R⁷)C(O)N(R⁷)₂, - OC(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)OR⁷, -OC(O)R⁷, -S(O)R⁷, and C_1-6 alkyl, wherein the C_1-6 alkyl, is optionally substituted with one or more substituents independently selected from halogen, -CN, - OH, -SH, -NO₂, and -NH₂;

R¹ is a linker selected from:

-O-, -S-, -N(R⁷)-, -C(O)-, -C(O)N(R⁷)-, -N(R⁷)C(O)-, -N(R⁷)C(O)N(R⁷)-, -OC(O)N(R⁷)-, - N(R⁷)C(O)O-, -C(O)O-, -OC(O)-, -S(O)-, -S(O)₂-, -OS(O)₂-, -OP(O)(OR⁷)O-, -SP(O)(OR⁷)O-, - OP(S)(OR⁷)O-, -OP(O)(SR⁷)O-, -OP(O)(OR⁷)S-, -OP(O)(O-)O-, -SP(O)(O-)O-, -OP(S)(O-)O-, - OP(O)(S-)O-, -OP(O)(O-)S-, -OP(O)(OR⁷)NR⁷-, -OP(O)(N(R⁷)₂)NR⁷-, -OP(OR⁷)O-, -OP(N(R⁷)₂)O-, - OP(OR⁷)N(R⁷)-, and -OPN(R⁷)₂NR⁷-; each R² is independently selected from:

C_1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, - OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂, -N(R⁷)C(O)R⁷, -N(R⁷)C(O)N(R⁷)₂, -OC(O)N(R⁷)₂, - N(R⁷)C(O)OR⁷, -C(O)OR⁷, -OC(O)R⁷, and -S(O)R⁷;

R³ and R⁴ are each independently selected from: -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂, -N(R⁷)C(O)R⁷, -N(R⁷)C(O)N(R⁷)₂, -OC(O)N(R⁷)₂, - N(R⁷)C(O)OR⁷, -C(O)OR⁷, -OC(O)R⁷, and -S(O)R⁷; each R⁵ is independently selected from:

-OC(O)R⁷, -OC(O)N(R⁷)₂, -N(R⁷)C(O)R⁷ -N(R⁷)C(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)R⁷, -C(O)OR⁷, and -C(O)N(R⁷)₂; each R⁶ is independently selected from: hydrogen; halogen, -CN, -NO₂, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂,-N(R⁷)C(O)R⁷ -N(R⁷)C(O)N(R⁷)₂, - OC(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)OR⁷, -OC(O)R⁷, and -S(O)R⁷; and

C_1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, - CN, -NO₂, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂, -N(R⁷)C(O)R⁷, -N(R⁷)C(O)N(R⁷)₂, - OC(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)OR⁷, -OC(O)R⁷, and -S(O)R⁷; each R⁷ is independently selected from: hydrogen;

C_1-6 alkyl, C_2-6 alkenyl, and C_2-6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O-C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkyl)₂, -NH(C_1-6 alkyl), C_3-10 carbocycle, and 3- to 10-membered heterocycle; and

C_3-10 carbocycle, and 3- to 10-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O- C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkyl)₂, -NH(C_1-6 alkyl), C_1-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_3-10 carbocycle, 3- to 10-membered heterocycle, and C 1 -6 haloalky 1.

(2) In some embodiments, each w is independently selected from any value from 1 to 10. In some embodiments, each w is independently selected from any value from 1 to 5. In some embodiments, each w is 1. In some embodiments, each v is independently selected from any value from 1 to 10. In some embodiments, each v is independently selected from any value from 1 to 5. In some embodiments, each v is 1. In some embodiments, n is selected from any value from 1 to 10. In some embodiments, n is selected from any value from 1 to 5. In some embodiments, n is 2. In some embodiments, m is selected from any value from 1 to 10. In some embodiments, m is selected from any value from 1 to 5. In some embodiments, m is selected from 1 and 2. In some embodiments, z is 3 and Y is C. In some embodiments, Q is selected from C5-6 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -NO₂, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -C(O)N(R⁷)₂, -N(R⁷)C(O)R⁷, -N(R⁷)C(O)N(R⁷)₂, -OC(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)OR⁷, - OC(O)R⁷, and -S(O)R⁷. In some embodiments, Q is selected from C5-6 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, and -NH₂. In some embodiments, Q is selected from phenyl and cyclohexyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, and -NH₂. In some embodiments, Q is selected from phenyl. In some embodiments, Q is selected from cyclohexyl. In some embodiments, R¹ is selected from -OP(O)(OR⁷)O-. -SP(O)(OR⁷)O- , -OP(S)(OR⁷)O-, -OP(O)(SR⁷)O-, -OP(O)(OR⁷)S-, -OP(O)(O-)O-, -SP(O)(O-)O-, -OP(S)(O-)O-, - OP(O)(S-)O-, -OP(O)(O-)S-, -OP(O)(OR⁷)NR⁷-, -OP(O)(N(R⁷)₂)NR⁷-, -OP(OR^O-, -OP(N(R⁷)₂)O-, - OP(OR⁷)N(R⁷)-, and -OPN(R⁷)₂NR⁷. In some embodiments, R¹ is selected from -OP(O)(OR⁷)O-, - SP(O)(OR⁷)O-, -OP(S)(OR⁷)O-, -OP(O)(SR⁷)O-, -OP(O)(OR⁷)S-, -OP(O)(O-)O-, -SP(O)(O-)O-, - OP(S)(O-)O-, -OP(O)(S-)O-, -OP(O)(O-)S-, and -OP(OR⁷)O-. In some embodiments, R¹ is selected from -OP(O)(OR⁷)O-, -OP(S)(OR⁷)O-, -OP(O)(O-)O-, -OP(S)(O-)O-, -OP(O)(S-)O-, and -OP(OR⁷)O- In some embodiments, R¹ is selected from -OP(O)(OR⁷)O- and -OP(OR⁷)O-. In some embodiments, R² is selected from C1-3 alkyl substituted with one or more substituents independently selected from halogen, -OR⁷, -OC(O)R⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, and -S(O)R⁷. In some embodiments, R² is selected from Ci -3 alkyl substituted with one or more substituents independently selected from -OR⁷, - OC(O)R⁷, -SR⁷, and -N(R⁷)₂. In some embodiments, R² is selected from C1-3 alkyl substituted with one or more substituents independently selected from -OR⁷ and -OC(O)R⁷. In some embodiments, R³ is selected from halogen, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, -OC(O)R⁷, and -S(O)R⁷. In some embodiments, R³ is selected from -OR⁷ -SR⁷, -OC(O)R⁷, and -N(R⁷)₂. In some embodiments, R³ is selected from - OR⁷ - and -OC(O)R⁷. In some embodiments, R⁴ is selected from halogen, -OR⁷, -SR⁷, -

N(R⁷)₂, -C(O)R⁷, -OC(O)R⁷, and -S(O)R⁷. In some embodiments, R⁴ is selected from -OR⁷ -SR⁷, - OC(O)R⁷, and -N(R⁷)₂.In some embodiments, R⁴ is selected from -OR⁷ - and -OC(O)R⁷. In some embodiments, R⁵ is selected from -OC(O)R⁷, -OC(O)N(R⁷)₂, -N(R⁷)C(O)R⁷ -N(R⁷)C(O)N(R⁷)₂, and - N(R⁷)C(O)OR⁷. In some embodiments, R⁵ is selected from -OC(O)R⁷ and -N(R⁷)C(O)R⁷. In some embodiments, each R⁷ is independently selected from: hydrogen; and C_1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O-C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkylX, -NH(C_1-6 alkyl), C_3-10 carbocycle, or 3- to 10- membered heterocycle. In some embodiments, each R⁷ is independently selected from C_1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O-C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkyl)₂, and -NH(C_1-6 alkyl). In some embodiments, each R⁷ is independently selected from C_1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, and -SH. In some embodiments, w is 1; v is 1; n is 2; m is 1 or 2; z is 3 and Y is C; Q is phenyl or cyclohexyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, and C_1-3 alkyl; R¹ is selected from -OP(O)(OR⁷)O-, -OP(S)(OR⁷)O-, -OP(O)(O-) O-, -OP(S)(Q- )O-, -OP(O)(S-)O-, and -OP(OR⁷)O-; R² is Ci alkyl substituted with -OH or -OC(O)CH₃; R³ is -OH or -OC(O)CH₃; R⁴ is -OH or -OC(O)CH₃; and R⁵ is -NH(O)CH₃. In some embodiments, the compound comprises:

[0148] In some embodiments, the oligonucleotide (J) is attached at a 5’ end or a 3’ end of the oligonucleotide. In some embodiments, the oligonucleotide comprises DNA. In some embodiments, the oligonucleotide comprises RNA. In some embodiments, the oligonucleotide comprises one or more modified intemucleoside linkages. In some embodiments, the one or more modified internucleoside linkages comprise alkylphosphonate, phosphorothioate, methylphosphonate, phosphor odi thioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, or carboxymethyl ester, or a combination thereof. In some embodiments, the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modified internucleoside linkages. In some embodiments, the compound binds to an asialoglycoprotein receptor. In some embodiments, the compound targets a hepatocyte.

[0149] Some embodiments include the following, where J is the oligonucleotide:

. J may include one or more additional phosphates, or one or more phosphorothioates linking to the oligonucleotide. J may include one or more additional phosphates linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide.

[0150] Some embodiments include the following, where J is the oligonucleotide:

[0151] Some embodiments include the following, where J is the oligonucleotide:

. J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.

[0152] Some embodiments include the following, where J is the oligonucleotide:

. The structure in this compound attached to the oligonucleotide (J) is an example of a GalNAc moiety. J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.

[0153] Some embodiments include the following, where J is the oligonucleotide:

[0154] Some embodiments include the following, where J is the oligonucleotide:

. J may include one or more additional phosphates, or one or more phosphorothioates linking to the oligonucleotide. J may include one or more additional phosphates linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. [0155] Some embodiments include the following, where J is the oligonucleotide:

J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.

[0156] Some embodiments include the following, where J is the oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL17,” and is an example of a GalNAc moiety. J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide. [0157] Some embodiments include the following, where the phosphate or “5 ’ ” indicates a connectiono the oligonucleotide:

[0158] Some embodiments include the following, where the phosphate or “5 ’ ” indicates a connection to the oligonucleotide:

[0159] Some embodiments include the following, where J is the oligonucleotide:

include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.

[0160] Some embodiments include the following, where J is the oligonucleotide:

. The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL1,” and is an example of a GalNAc moiety. J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.

[0161] Some embodiments include the following, where J is the oligonucleotide:

Some embodiments include the following, where J is the oligonucleotide:

[0162] Some embodiments include the following, where J is the oligonucleotide:

[0163] Some embodiments include the following, where J is the oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL17,” and is an example of a GalNAc moiety. J may include one or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a Disclosed herein, in some embodiments, are compositions comprising an oligonucleotide that inhibits the expression of a target gene, wherein the oligonucleotide comprises a GalNAc moiety. The GalNAc moiety may be included in any formula, structure, or GalNAc moiety shown below. In some embodiments, described herein is a compound (e.g., oligonucleotide) represented by Formula (III), (IV), or (V):

or a salt

thereof, wherein

J is an oligonucleotide; each w is independently selected from any value from 0 to 20; v is independently selected from any value from 0 to 20; each n is selected from any value from 0 to 20; each m is selected from any value from 0 to 20; each p is selected from any value from 0 to 1; each w is selected from any value from 0 to 20; t is selected from any value from 0 to 1 ; x is selected from any value from 0 to 1 ; r is selected from any value from 0 to 20; u is selected from any value from 0 to 20;

Q is selected from: C_3-20 cyclic, heterocyclic or acyclic linker optionally substituted with one or more substituents independently selected from halogen, -CN, -NO₂, -OR⁷, -SR⁷, -N(R⁷)₂, -C(O)R⁷, - C(O)N(R⁷)₂, -N(R⁷)C(O)R⁷, -N(R⁷)C(O)N(R⁷)₂, -OC(O)N(R⁷)₂, -N(R⁷)C(O)OR⁷, -C(O)OR⁷, - OC(O)R⁷, -S(O)R⁷, and C_1-6 alkyl, wherein the C_1-6 alkyl, is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, and -NH₂;

R¹ is a linker selected from: -O-, -S-, -N(R⁷)-, -C(O)-, -C(O)N(R⁷)-, -N(R⁷)C(O)-. -N(R⁷)C(O)N(R⁷)-, -OC(O)N(R⁷)-, -N(R⁷)C(O)O-, -C(O)O-, -OC(O)-, -S(O)-, -S(O)₂-, -OS(O)₂-, -OP(O)(OR⁷)O-, - SP(O)(OR⁷)O-, -OP(S)(OR⁷)O-, -OP(O)(SR⁷)O-, -OP(O)(OR⁷)S-, -OP(O)(O-)O-, -SP(O)(O-)O-, - OP(S)(O-)O-, -OP(O)(S-)O-, -OP(O)(O-)S-, -OP(O)(OR⁷)NR⁷-, -OP(O)(N(R⁷)₂)NR⁷-, -OP(OR⁷)O-, - OP(N(R⁷)₂)O-, -OP(OR⁷)N(R⁷)-, and -OPN(R⁷)₂NR⁷-; each R⁷ is independently selected from: hydrogen, C_1-6 alkyl, C_2-6 alkenyl, and C_2-6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, - CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O-C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkyl)₂, -NH(C_1-6 alkyl), C_3-10 carbocycle, and 3- to 10-membered heterocycle, C_3-10 carbocycle, and 3- to 10-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO₂, -NH₂, =O, =S, -O-C_1-6 alkyl, -S-C_1-6 alkyl, -N(C_1-6 alkyl)₂, -NH(C_1-6 alkyl), C_1-6 alkyl, C₂-6 alkenyl, C₂-6 alkynyl, C_3-10 carbocycle, 3- to 10-membered heterocycle, and C_1-6 haloalkyl.

[0164] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[0165] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “L96,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphor othi oates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphor othi oates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0166] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[0167] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “NAG37,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphor othi oates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothi oates linking to the oligonucleotide. J in some instances comprises a phosphorothi oate linking to the oligonucleotide.

[0168] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[0169] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “GluGalNAc,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0170] Provided herein are sugar moieties comprising the following structure, where J and K are independently H, a GalNAc moiety or oligonucleotides:

[0171] The structures in these compounds in some instances are attached to the oligonucleotide (J or K) and referred to as “ademA GalNAc, ademG GalNAc, ademC GalNAc, or ademU GalNAc” depending on the base used in the nucleotide. In some instances, 2-4 GalNAc moieties are attached to the oligonucleotide. The placement of the GalNAc moieties in some instances is at the 3 or 5’ ends (J or K = H) or internal (J and K are oligonucleotides) of the oligonucleotide strand. J and K may in some instances comprises one or more phosphates or phosphorothi oates linking to the oligonucleotide. J and K in some instances comprises one or more phosphates linking to the oligonucleotide. J and K in some instances comprises a phosphate linking to the oligonucleotide. J and K in some instances comprises one or more phosphorothi oates linking to the oligonucleotide. J and K in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0172] Provided herein are sugar moieties comprising the following structure, where R is an oligonucleotide:

[0173] The structure in this compound attached to the oligonucleotide (R) in some instances is referred to as H1, H2, H3, H4, H5, H6, H7, or H9, and are examples of GalNAc moi eties. R in some instances comprises one or more phosphates or phosphorothi oates linking to the oligonucleotide. R in some instances comprises one or more phosphates linking to the oligonucleotide. R in some instances comprises a phosphate linking to the oligonucleotide. R in some instances comprises one or more phosphorothioates linking to the oligonucleotide. R in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0174] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) may be referred to as “K2GalNAc,' and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide. [0175] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide and X is S or O:

. The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “ST23,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide. [0176] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “GalNAc23,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide. [0177] Provided herein are sugar moieties comprising the following structure, where J or K comprises an oligonucleotide:

[0178] The structures in these compounds in some instances are attached to the oligonucleotide (J or K), referred to as “PyrGalNAc”, “PipGalNAc” and “TEG-GalNAc” are examples of GalNAc moieties. In some instances, 2-4 GalNAc moieties are attached oligonucleotide. The placement of the GalNAc moieties may be at the 3 or 5’ ends (J or K = H) or internal (J and K are oligonucleotides) of the oligonucleotide strand. J and K in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J and K in some instances comprises one or more phosphates linking to the oligonucleotide. J and K in some instances comprises a phosphate linking to the oligonucleotide. J and K in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J and K in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0179] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[0180] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “THA,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphor othi oates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphor othi oates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0181] Provided herein are sugar moieties comprising the following structure, where Nu is an oligonucleotide:

[0182] The structure in this compound attached to the oligonucleotide (Nu) in some instances is referred to as “L-9” and is an example of a GalNAc moiety. Nu in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. Nu in some instances comprises one or more phosphates linking to the oligonucleotide. Nu in some instances comprises a phosphate linking to the oligonucleotide. Nu in some instances comprises one or more phosphorothioates linking to the oligonucleotide. Nu in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0183] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[00159] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “Sirius GalNAc,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[00160] Provided herein are sugar moieties comprising the following structures, where J is an oligonucleotide:

[0184] The structures in this compound attached to the oligonucleotide (J) in some instances are referred to as GLS-5 and GLS-15 and are examples of GalNAc moieties. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0185] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

[0186] The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “Olix GalNAc,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0187] Provided herein are sugar moieties comprising the following structure, where J and J’ is an oligonucleotide or a GalNAc moiety:

[0188] The structure in this compound attached to the oligonucleotide or a GalNAc moiety (J or J’) in some instances is referred to as “GalNAc Gib,” and is an example of a GalNAc moiety. J or J’ in some instances comprises one or more phosphates or phosphorothi oates linking to the oligonucleotide. J or J’ in some instances comprises one or more phosphates linking to the oligonucleotide. J or J’ in some instances comprises a phosphate linking to the oligonucleotide. J or J’ in some instances comprises one or more phosphorothi oates linking to the oligonucleotide. J or J’ in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0189] Provided herein are sugar moieties comprising the following structure, where B is a nucleic acid base, and J and J’ is an oligonucleotide or a GalNAc moiety:

[0190] The structure in this compound attached to the oligonucleotide or a GalNAc moiety (J or J’) in some instances is referred to as “lgT3,” and is an example of a GalNAc moiety. J or J’ in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J or J’ in some instances comprises one or more phosphates linking to the oligonucleotide. J or J’ in some instances comprises a phosphate linking to the oligonucleotide. J or J’ in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J or J’ in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0191] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide and X is an optional linker:

The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “5gn2c6,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide. X is a carbon or heteroatom linker to J. In some instances, the heteroatom in linker X is an N or O. [0192] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “[Gal-6]s[Gal-6]s[Gal-6],” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

[0193] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “Janssen,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide. [0194] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:

The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “Arbutus,” and is an example of a GalNAc moiety. J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. J in some instances comprises one or more phosphates linking to the oligonucleotide. J in some instances comprises a phosphate linking to the oligonucleotide. J in some instances comprises one or more phosphorothioates linking to the oligonucleotide. J in some instances comprises a phosphorothioate linking to the oligonucleotide.

3. Modified siRNAs

[0195] In some embodiments, the composition comprises an oligonucleotide that inhibits or reduces the expression of target nucleic acid, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand comprises modification

[0196] In all the above modification pattern, wherever they occur, “Nf” is a 2’ -fluoro-modified nucleoside, “n” is a 2’ -O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, “dN” is a 2’-deoxy-modified nucleoside or a 2’ -deoxy nucleoside, “nm” is a 2’-O-methoxyethyl modified nucleoside, “i” is an inosine, “ni” is a 2’ -O-methyl inosine nucleoside, N(C16) is 2’-O- hexadecate modification andN comprises one or more nucleosides. In some modifications N(C16) is a 2'-O-hexadecyl adenylate.

modification pattern, wherever they occur, “Nf” is a 2’ -fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, “dN” is a 2’ -deoxymodified nucleoside or a 2’ -deoxy nucleoside, “nm” is a 2’ -O-methoxy ethyl modified nucleoside, “i” is an inosine, “ni” is a2’-O-methyl inosine nucleoside, N(C16) is 2’-O-hexadecate modification, VP is a 5'- vinyl phosphonate, “[NUNA]” is an unlocked nucleic acid, “[UNA]” is an unlocked nucleic acid and N comprises one or more nucleosides. In some modifications N(C16) is a 2'-O-hexadecyl adenylate.

[0198] In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand comprises pattern IS and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45 AS, 46AS, 47AS, 48AS, 49AS, 50As, 51AS, 52AS, 53AS, 54AS 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 2S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49 AS, 5OAs, 51AS, 52AS, 53AS, 54AS 55AS, 56AS, 57AS, 58AS, 59AS, 60 AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, 69AS, 70AS, 71 AS, or72AS . In some embodiments, the sense strand comprises pattern 3S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS,

47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 4S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 5S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 6S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS,

40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS,

54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 7S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 8S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS,

45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 9S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 10S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 11S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9 AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 12S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

26AS, 27 AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 13S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 14S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 15S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53 AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 16S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 17S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 18S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43AS, 44AS, 45 AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 19S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 20S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 21S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43 AS, 44AS, 45 AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55 AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 22S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 23 S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 24S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 25S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 26S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 27S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 28S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40 AS, 41 AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 29S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 30S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7 AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15 AS. 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29 AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 3 IS and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 32S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 33S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 34S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 35S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 36S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 37S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 38S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 39S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 40S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 41S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 42S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60 AS, 61AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 43 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 44S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 45 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53 AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 46S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 47S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 48S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 49S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 50S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 51 S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises patern 52S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises patern 53S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises patern 54S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, I 5AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises patern 55S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 56S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 57S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 58S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45 AS, 46AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 59S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, or 40 AS. In some embodiments, the sense strand comprises pattern 60S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS,

24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS,

38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS,

52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS,

66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 61S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 62S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises patern 63S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises patern 64S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS,

31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS,

59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 65S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises patern 66S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises patern 67S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises patern 68S and the antisense strand comprises patern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 69S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 70S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 71 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 72S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 73S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 74S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 75S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 76S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 77S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40 AS, 41 AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 78S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 79S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 80S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 81S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS, In some embodiments, the sense strand comprises pattern 82S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 83S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 84S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 85 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 86S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15 AS, 16AS, 17 AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 87S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15 AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 88S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 89S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 90S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63 AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 91 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21 AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 92S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 93 S and the antisense strand comprises pattern 1AS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 94S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 95S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 96S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 97S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 98S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 99S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 100S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 101 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 102S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 103S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 104S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 105S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 106S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS,

30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS,

44AS, 45 AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53 AS, 54AS, 55 AS, 56AS, 57AS,

58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 107S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 108S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 109S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 110S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20 AS, 21AS, 22 AS, 23AS, 24AS, 25 AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 11 IS and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS,

68AS, or 69AS. In some embodiments, the sense strand comprises pattern 112S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 113S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 114S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25 AS, 26AS, 27AS, 28AS, 29AS,

58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65 AS, 66AS, 67AS, 68AS, or 69AS. In some embodiments, the sense strand comprises pattern 115S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33 AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41 AS, 42As, 43 AS, 44AS, 45AS, 46AS, 47 AS, 48AS, 49 AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61 AS, 62AS, 63 AS, 64AS, 65 AS, 66AS, 67 AS, 68AS, or 69 AS. In some embodiments, the sense strand comprises pattern 116S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13 AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS,

61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67AS, 68AS, or 69AS.

[0199] In some embodiments, the sense strand comprises pattern 117S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. [0200] In some embodiments, the sense strand comprises pattern 118S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0201] In some embodiments, the sense strand comprises pattern 119S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0202] In some embodiments, the sense strand comprises pattern 120S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0203] In some embodiments, the sense strand comprises pattern 12 IS and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0204] In some embodiments, the sense strand comprises pattern 122S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0205] In some embodiments, the sense strand comprises pattern 123S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0206] In some embodiments, the sense strand comprises pattern 124S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0207] In some embodiments, the sense strand comprises pattern 125S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0208] In some embodiments, the sense strand comprises pattern 126S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0209] In some embodiments, the sense strand comprises pattern 127S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0210] In some embodiments, the sense strand comprises pattern 128S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0211] In some embodiments, the sense strand comprises pattern 129S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0212] In some embodiments, the sense strand comprises pattern 130S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0213] In some embodiments, the sense strand comprises pattern 13 IS and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40 AS, 41 AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0214] In some embodiments, the sense strand comprises pattern 132S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40 AS, 41 AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0215] In some embodiments, the sense strand comprises pattern 133S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29 AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40 AS, 41 AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0216] In some embodiments, the sense strand comprises pattern 134S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0217] In some embodiments, the sense strand comprises pattern 135S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0218] In some embodiments, the sense strand comprises pattern 136S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0219] In some embodiments, the sense strand comprises pattern 137S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0220] In some embodiments, the sense strand comprises pattern 138S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0221] In some embodiments, the sense strand comprises pattern 139S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0222] In some embodiments, the sense strand comprises pattern 140S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0223] In some embodiments, the sense strand comprises pattern 141 S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0224] In some embodiments, the sense strand comprises pattern 142S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0225] In some embodiments, the sense strand comprises pattern 143S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0226] In some embodiments, the sense strand comprises pattern 144S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0227] In some embodiments, the sense strand comprises pattern 145S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0228] In some embodiments, the sense strand comprises pattern 146S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26 AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0229] In some embodiments, the sense strand comprises pattern 147S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0230] In some embodiments, the sense strand comprises pattern 148S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0231] In some embodiments, the sense strand comprises pattern 149S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0232] In some embodiments, the sense strand comprises pattern 150S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68 AS, or 69AS [0233] In some embodiments, the sense strand comprises pattern 15 IS and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0234] In some embodiments, the sense strand comprises pattern 152S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0235] In some embodiments, the sense strand comprises pattern 153S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS [0236] In some embodiments, the sense strand comprises pattern 154S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. [0237] In some embodiments, the sense strand comprises pattern 155S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. [0238] In some embodiments, the sense strand comprises pattern 156S and the antisense strand comprises pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23AS, 24AS, 25AS, 26AS, 27AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42As, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS, 50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS, 64AS, 65AS, 66AS, 67 AS, 68AS, or 69AS. [0239] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S,

108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S,

124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 1AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S.

131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S,

147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 2AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S,

9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S, 104S, 105S, 106S,

107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S,

123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S,

139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S,

155S, or 156S, and the antisense strand comprises pattern 3AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S,

38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S,

58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S,

78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S,

98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 4AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S,

27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S,

47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S,

67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S,

87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S,

105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S,

121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S,

137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S,

153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 5AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S,

36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S,

56S, 57S, 58S, 59S, 60S, 61 S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S,

76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S,

96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 6AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51 S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S,

103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S,

119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S,

135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S,

151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 7AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S,

53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S,

73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S,

93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111s, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S,

126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 8AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S , 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S. 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S,

42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S,

62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S,

82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S,

101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S,

117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S,

133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S,

149S, 150S, 151S, 152S, 153S, 154S, 155S, or l56S, and the antisense strand comprises pattern 9AS.

In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S,

31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S,

51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S,

71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S,

91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S,

124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 10AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31 S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51 S, 52S, 53S, 54S, 55S, 56S, 57S, 58S,

59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S,

79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S.

147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 11AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S,

48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S,

68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S,

88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S, 104S, 105S,

106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S,

122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S,

138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S,

154S, 155S, or 156S, and the antisense strand comprises pattern 12AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S,

37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S,

57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S,

77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S,

97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 13AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S,

45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S,

65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S,

85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S,

104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S,

120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S,

136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S,

152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 14AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 15 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S,

41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S,

61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S,

81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S,

100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S,

116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S,

132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S,

148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern

16AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S,

50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S,

70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S,

90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 17AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, 156S, 157S, 158S, 159S, 160S, 161S, 162S, 163S, or 164S and the antisense strand comprises pattern 18AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S,

78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S , 97S,

98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 19AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S,

121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S,

153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 20AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S,

56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S,

96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 21AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S,

43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S,

63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S,

83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S,

102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 22AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S,

32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S,

52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S,

72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S,

92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 23AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 24AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S,

122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S,

154S, 155S, or 156S, and the antisense strand comprises pattern 25 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S,

77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S , 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S , 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 26 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S,

85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S,

120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S,

152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 27 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11 S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, I l ls, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 28 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S,

21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S,

81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S,

29AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S,108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 30AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 31AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S,

154S, 155S, or 156S, and the antisense strand comprises pattern 32AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S,

97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, I 30S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 33AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51 S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S,

152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 34AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11 S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 35 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S,

36AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 37AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 38AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S,

154S, 155S, or 156S, and the antisense strand comprises pattern 39AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S,

97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 40AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S,

152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 41AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 42AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S,

43AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 44AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 45AS.In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S,

155S, or 156S, and the antisense strand comprises pattern 46AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S,

98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 47 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S,

153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 48AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S,

96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 49 AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S,

83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S,

102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S,

118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S,

134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S,

150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 50AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S,

92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, I 30S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 51AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, I 30S.

147S, 148S, 149S, 150S, 151 S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 52AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S,

88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S,

154S, 155S, or 156S, and the antisense strand comprises pattern 53AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S,

97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S. 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 54AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S,

152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 55AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11 S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 56AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S,

61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S,

132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145 S, 146S, 147S,

57AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S, and the antisense strand comprises pattern 58AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S,

99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S, 130S,

147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, 156S, 157S, 158S, 159S, 160S, 161S, 162S,

163S, or 164S and the antisense strand comprises pattern 59AS. In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S,

98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S, 126S, 127S, 128S, 129S,

130S, 131S, 132S, 133S, 134S, 135S, 136S, 137S, 138S, 139S, 140S, 141S, 142S, 143S, 144S, 145S,

146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, 156S, 157S, 158S, 159S, 160S, 161S,

162S, 163S, or 164S and the antisense strand comprises pattern 60AS.

[0240] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S,

90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101 S, 102S, 103S, 104S, 105S, 106S, 107S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S,

156S, 157S, 158S, 159S, 160S, 161S, 162S, 163S, or 164S and the antisense strand comprises pattern

61AS.

[0241] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S,

62AS.

[0242] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

63 AS.

[0243] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

64AS.

[0244] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

65 AS.

[0245] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

66AS.

[0246] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

140S, 141S, 142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, 156S, 157S, 158S, 159S, 160S, 161S, 162S, 163S, or 164S and the antisense strand comprises pattern 67 AS.

[0247] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

68AS.

[0248] In some embodiments, the sense strand comprises pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S,

69AS.

[0249] In some embodiments, the sense strand comprises any one of modification patters 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, or 9S. In some embodiments, the sense strand comprises any one of modification patters 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S, 110S, 111S, 112S, 113S, 114S, 115S,

148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S. In some embodiments, the sense strand comprises modification pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, or 8AS. In some embodiments, the antisense strand comprises modification pattern 1AS, 2AS, 3AS, 4AS, 5 AS, 6AS, 7 AS, 8AS, 9AS, 10AS, 11AS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21AS, 22AS, 23 AS, 24AS, 25 AS, 26 AS, 27 AS, 28AS, 29AS, 30AS, 31AS, 32AS, 33AS, 34AS, 35 AS,

36AS, 37AS, 38AS, 39AS, 40AS, 41AS, 42AS, 43AS, 44AS, 45AS, 46AS, 47AS, 48AS, 49AS,

50AS, 51AS, 52AS, 53AS, 54AS, 55AS, 56AS, 57AS, 58AS, 59AS, 60AS, 61AS, 62AS, 63AS,

64AS, 65 AS, 66AS, 67 AS, 68AS, or 69AS. In some embodiments, the antisense strand comprises modification pattern 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, or 8AS. In some embodiments, the antisense strand comprises modification pattern 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11 S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 69S, 70S, 71S, 72S, 73S, 74S, 75S, 76S, 77S, 78S, 79S, 80S, 81S, 82S, 83S, 84S, 85S, 86S, 87S, 88S, 89S, 90S, 91S, 92S, 93S, 94S, 95S, 96S, 97S, 98S, 99S, 100S, 101S, 102S, 103S, 104S, 105S, 106S, 107S, 108S, 109S,

110S, I l ls, 112S, 113S, 114S, 115S, 116S, 117S, 118S, 119S, 120S, 121S, 122S, 123S, 124S, 125S,

142S, 143S, 144S, 145S, 146S, 147S, 148S, 149S, 150S, 151S, 152S, 153S, 154S, 155S, or 156S. In some embodiments, the sense strand or the antisense strand comprises modification pattern ASO1. [0250] In some embodiments, purines of the sense strand comprise 2’ -fluoro modified purines. In some embodiments, purines of the sense strand comprise 2’ -O-methyl modified purines. In some embodiments, purines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all purines of the sense strand comprise 2’ -fluoro modified purines. In some embodiments, all purines of the sense strand comprise 2’ -O-methyl modified purines. In some embodiments, all purines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines.

[0251] In some embodiments, pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines.

[0252] In some embodiments, purines of the sense strand comprise 2’ -fluoro modified purines, and pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the sense strand comprise 2’ -O-methyl modified purines, and pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the sense strand comprise 2’ -fluoro modified purines, and pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the sense strand comprise 2’ -O-methyl modified purines, and pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines, and purines of the sense strand comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines. In some embodiments, pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines, and purines of the sense strand comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines. In some embodiments, pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines, and purines of the sense strand comprise 2’-O- methyl modified purines. In some embodiments, pyrimidines of the sense strand comprise 2’ -O- methyl modified pyrimidines, and purines of the sense strand comprise 2’ -fluoro modified purines. [0253] In some embodiments, all purines of the sense strand comprise 2’ -fluoro modified purines, and all pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the sense strand comprise 2’ -O-methyl modified purines, and all pyrimidines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the sense strand comprise 2’ -fluoro modified purines, and all pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the sense strand comprise 2’ -O-methyl modified purines, and all pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines, and all purines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines, and all purines of the sense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -fluoro modified pyrimidines, and all purines of the sense strand comprise 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the sense strand comprise 2’ -O-methyl modified pyrimidines, and all purines of the sense strand comprise 2’ -fluoro modified purines.

[0254] In some embodiments, purines of the antisense strand comprise 2’ -fluoro modified purines. In some embodiments, purines of the antisense strand comprise 2’ -O-methyl modified purines. In some embodiments, purines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, all purines of the antisense strand comprise 2’ -fluoro modified purines. In some embodiments, all purines of the antisense strand comprise 2’ -O-methyl modified purines. In some embodiments, all purines of the antisense strand comprise a mixture of 2’ - fluoro and 2’ -O-methyl modified purines.

[0255] In some embodiments, pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines.

[0256] In some embodiments, purines of the antisense strand comprise 2’ -fluoro modified purines, and pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the antisense strand comprise 2’ -O-methyl modified purines, and pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the antisense strand comprise 2’ -fluoro modified purines, and pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, purines of the antisense strand comprise 2’ -O-methyl modified purines, and pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines, and purines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines, and purines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines, and purines of the antisense strand comprise 2’ -O-methyl modified purines. In some embodiments, pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines, and purines of the antisense strand comprise 2’ -fluoro modified purines.

[0257] In some embodiments, all purines of the antisense strand comprise 2’ -fluoro modified purines, and all pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the antisense strand comprise 2’ -O-methyl modified purines, and all pyrimidines of the antisense strand comprise a mixture of 2’ -fluoro and 2’- O-methyl modified pyrimidines. In some embodiments, all purines of the antisense strand comprise 2’-fluoro modified purines, and all pyrimidines of the antisense strand comprise 2’ -O-methyl modified pyrimidines. In some embodiments, all purines of the antisense strand comprise 2’ -O-methyl modified purines, and all pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines, and all purines of the antisense strand comprise a mixture of 2’ -fluoro and 2’- O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the antisense strand comprise a mixture of 2’- fluoro and 2’ -O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’ -fluoro modified pyrimidines, and all purines of the antisense strand comprise 2’ - O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the antisense strand comprise 2’ -fluoro modified purines.

[0258] In some embodiments, the siRNA comprises a sense strand, an antisense strand, and a lipid moiety connected to an end of the sense or antisense strand; wherein the lipid moiety comprises a phenyl or cyclohexanyl linker, wherein the linker is connected to a lipid and to the end of the sense or antisense strand. In some embodiments, any one of the following is true with regard to the sense strand: (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O- methyl, and 2’-O-methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O-methyl modified purines and all pyrimidines comprise (vi) all pyrimidines of the sense strand comprise a mixture of 2’- fluoro and 2’ -O-methoxyethyl modified pyrimidines; or (ii) amixture of 2’-fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (c) all purines comprise 2'-O-methoxyethyl modified purines and all pyrimidines comprise (i) a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; or (ii) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’ -O-methoxyethyl modified pyrimidines; (d) all purines comprise a mixture of 2’ -fluoro and 2'-O-methyl modified purines and all pyrimidines comprise (i) 2’ -O-methoxyethyl modified pyrimidines; (ii) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; (iii) a mixture of 2’-fluoro and 2’ -O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’ -O-methoxyethyl modified pyrimidines; (e) all purines comprise a mixture of 2’ -fluoro and 2'-O-methoxy ethyl modified purines and all pyrimidines of the sense strand comprise (i) 2’-O-methyl modified pyrimidines; (ii) a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; (iii) a mixture of 2’-O-methyl and 2’-O- methoxy ethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’-O- methoxyethyl modified pyrimidines; (f) all purines comprise a mixture of 2'-O-methyl and 2'-O- methoxy ethyl modified purines and all pyrimidines comprise (i) 2’-fluoro modified pyrimidines; (ii) a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; (iii) a mixture of 2’ -fluoro and 2’-O- methoxy ethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’-O- methoxy ethyl modified pyrimidines; or (g) all purines comprise a mixture of 2’-fluoro, 2’-O-methyl, and 2’ -O-methoxyethyl modified purines and all pyrimidines comprise (i) 2’-fluoro modified pyrimidines; (ii) 2’-O-methyl modified pyrimidines; (iii) 2’ -O-methoxyethyl modified pyrimidines; (iv) a mixture of 2’ -fluoro and 2’-O-meihyl modified pyrimidines; (v) a mixture of 2’-O-methyl and 2’-O-methoxy ethyl modified pyrimidines; (vi) a mixture of 2’-fluoro and 2’ -O-methoxyethyl modified pyrimidines; or (vii) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’ -O-methoxyethyl modified pyrimidines. In some embodiments, any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’- fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise 2’- fluoro modified purines. In some embodiments, the siRNA comprises comprising a sense strand and an antisense strand; wherein the antisense strand comprises a 5’ end comprising a vinyl phosphonate and 2 phosphorothioate linkages, and a 3’ end comprising 2 phosphorothioate linkages; wherein the sense strand comprises (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’- fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O- methyl modified purines and all pyrimidines comprise (vi) all pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methoxyethyl modified pyrimidines; or (ii) amixture of 2’- fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (c) all purines comprise 2'-O- methoxy ethyl modified purines and all pyrimidines comprise (i) a mixture of 2’-fluoro and 2’-O- methyl modified pyrimidines; or (ii) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (d) all purines comprise a mixture of 2’-fluoro and 2'-O-methyl modified purines and all pyrimidines comprise (i) 2’-O-methoxyethyl modified pyrimidines; (ii) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; (iii) a mixture of 2’-fluoro and 2’-O- methoxy ethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’-O- methoxyethyl modified pyrimidines; (e) all purines comprise a mixture of 2’-fluoro and 2'-O- methoxyethyl modified purines and all pyrimidines of the sense strand comprise (i) 2’ -O-methyl modified pyrimidines; (ii) a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines; (iii) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’-O-methyl, and 2’-O-methoxyeihyl modified pyrimidines; (f) all purines comprise a mixture of 2'- O-methyl and 2'-O-methoxy ethyl modified purines and all pyrimidines comprise (i) 2’ -fluoro modified pyrimidines; (ii) a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines; (iii) a mixture of 2’-fluoro and 2’-O-methoxyethyl modified pyrimidines; or (iv) amixture of 2’-fluoro, 2’- O-methyl, and 2’-O-methoxyeihyl modified pyrimidines; or (g) all purines comprise a mixture of 2’- fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified purines and all pyrimidines comprise (i) 2’- fluoro modified pyrimidines; (ii) 2’ -O-methyl modified pyrimidines; (iii) 2’-O-methoxyethyl modified pyrimidines; (iv) a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines; (v) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; (vi) amixture of 2’ -fluoro and 2’-O-methoxyethyl modified pyrimidines; or (vii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O- methoxy ethyl modified pyrimidines; and wherein any one of the following is true with regard to the antisense strand: all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise 2’- fluoro modified pyrimidines, all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines, all pyrimidines comprise 2’-O- methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines, or all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise 2’ -fluoro modified purines.

[0259] In some embodiments, any one of the following is true with regard to the sense strand: (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O- methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O-methyl modified purines and all pyrimidines comprise (vi) all pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’- O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O- methoxy ethyl modified pyrimidines; (c) all purines comprise 2'-O-methoxyethyl modified purines and all pyrimidines comprise (i) a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines; or (ii) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (d) all purines comprise a mixture of 2’ -fluoro and 2'-O-methyl modified purines and all pyrimidines comprise (i) 2’- O-methoxyethyl modified pyrimidines; (ii) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; (iii) a mixture of 2’ -fluoro and 2’-O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’ -O-meihoxy ethyl modified pyrimidines; (e) all purines comprise a mixture of 2’ -fluoro and 2'-O-methoxy ethyl modified purines and all pyrimidines of the sense strand comprise (i) 2’ -O-methyl modified pyrimidines; (ii) a mixture of 2’ -fluoro and 2’-O- methyl modified pyrimidines; (iii) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (1) all purines comprise a mixture of 2'-O-methyl and 2'-O-methoxyethyl modified purines and all pyrimidines comprise (i) 2’-fluoro modified pyrimidines; (ii) a mixture of 2’-fluoro and 2’ -O-methyl modified pyrimidines; (iii) a mixture of 2’ -fluoro and 2’-O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified pyrimidines; or (g) all purines comprise a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified purines and all pyrimidines comprise (i) 2’ -fluoro modified pyrimidines; (ii) 2’ -O-methyl modified pyrimidines; (iii) 2’-O-methoxyethyl modified pyrimidines; (iv) a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; (v) a mixture of 2’ -O-methyl and 2’-O-methoxyethyl modified pyrimidines; (vi) a mixture of 2’-fluoro and 2’-O-methoxyethyl modified pyrimidines; or (vii) a mixture of 2’ -fluoro, 2’ -O-methyl, and 2’-O-methoxyethyl modified pyrimidines. In some embodiments, a deoxy nucleoside may be included in the sense strand. In some embodiments, the sense strand includes the deoxy nucleoside. The deoxy nucleoside may be at nucleoside position 9 of the sense strand. In some embodiments, the sense strand does not include a deoxy nucleoside. The deoxy nucleoside of the sense strand may be otherwise unmodified.

[0260] In some embodiments, the oligonucleotide comprises a duplex consisting of 21 nucleotide single strands with base pairing between 19 of the base pairs. In some embodiments, the duplex comprises single-stranded 2 nucleotide overhangs are at the 3’ ends of each strand. One strand (antisense strand) is complementary to a GPAM mRNA. Each end of the antisense strand has one to two phosphorothioate bonds. The 5’ end has an optional phosphate mimic such as a vinyl phosphonate. In some embodiments, the oligonucleotide is used to knock down a GPAM mRNA or a target protein. In some embodiments, the sense strand has the same sequence as the GPAM mRNA. In some embodiments, there are 1-2 phosphorothioates at the 3’ end. In some embodiments, there are 1 or no phosphorothioates at the 5’ end. In some embodiments, there is a hydrophobic conjugate of 12 to 25 carbons attached at the 5’ end via a phosphodi ester bond.

[0261] Disclosed herein, in some embodiments, are modified oligonucleotides. The modified oligonucleotide may be an siRNA that includes modifications to the ribose rings, and phosphate linkages. The modifications may be in particular patterns that maximize cell delivery, stability, and efficiency. The siRNA may also include a vinyl phosphonate and a hydrophobic group. These modifications may aid in delivery to a cell or tissue within a subject. The modified oligonucleotide may be used in a method such as a treatment method or a method of reducing gene expression. [0262] In some embodiments, the oligonucleotide comprises a duplex consisting of 21 nucleotide single strands with base pairing between 19 of the base pairs. In some embodiments, the duplex comprises single-stranded 2 nucleotide overhangs are at the 3’ ends of each strand. One strand (antisense strand) is complementary to a GPAM mRNA. Each end of the antisense strand has one to two phosphorothioate bonds. The 5’ end has an optional phosphate mimic such as a vinyl phosphonate. In some embodiments, the oligonucleotide is used to knock down a GPAM mRNA or a target protein. In some embodiments, the sense strand has the same sequence as the GPAM mRNA. In some embodiments, there are 1-2 phosphorothioates at the 3’ end. In some embodiments, there are 1 or no phosphorothioates at the 5’ end. In some embodiments, there is a hydrophobic conjugate of 12 to 25 carbons attached at the 5’ end via a phosphodi ester bond.

[0263] In some cases, the sense strand of any of the siRNAs comprises siRNA with a particular modification pattern. In some embodiments of the modification pattern, position 9 counting from the 5’ end of the sense strand may have a 2’F modification. In some embodiments, when position 9 of the sense strand is a pyrimidine, then all purines in the sense strand have a 2’ OMe modification. In some embodiments, when position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in the sense strand. In some embodiments, when position 9 and only one other base between positions 5 and 11 of the sense strand are pyrimidines, then both of these pyrimidines are the only two positions with a 2’F modification in the sense strand. In some embodiments, when position 9 and only two other bases between positions 5 and 11 of the sense strand are pyrimidines, and those two other pyrimidines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. In some embodiments, when there are more than 2 pyrimidines between positions 5 and 11 of the sense strand, then all combinations of pyrimidines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that the sense strand does not have three 2’F modifications in a row. In some cases, the sense strand of any of the siRNAs comprises a modification pattern which conforms to any or all of these sense strand rules.

[0264] In some embodiments, when position 9 of the sense strand is a purine, then all purines in the sense strand have a 2’OMe modification. In some embodiments, when position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in the sense strand. In some embodiments, when position 9 and only one other base between positions 5 and 11 of the sense strand are purines, then both of these purines are the only two positions with a 2’F modification in the sense strand. In some embodiments, when position 9 and only two other bases between positions 5 and 11 of the sense strand are purines, and those two other purines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. In some embodiments, when there are more than 2 purines between positions 5 and 11 of the sense strand, thai all combinations of purines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that the sense strand does not have three 2’F modifications in a row. In some cases, the sense strand of any of the siRNAs comprises a modification pattern which conforms to any or all of these sense strand rules.

[0265] In some embodiments, the siRNA comprises a sense strand, an antisense strand, and a lipid moiety connected to an end of the sense or antisense strand; wherein the lipid moiety comprises a phenyl or cyclohexanyl linker, wherein the linker is connected to a lipid and to the end of the sense or antisense strand. In some embodiments, any one of the following is true with regard to the sense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’- fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise 2’-O-methyl modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines. In some embodiments, any one of the following is true with regard to the antisense strand: all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’- fluoro modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O- methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise 2’ -fluoro modified purines. In some embodiments, the siRNA comprises comprising a sense strand and an antisense strand; wherein the antisense strand comprises a 5’ end comprising a vinyl phosphonate and 2 phosphorothioate linkages, and a 3’ end comprising 2 phosphorothioate linkages; wherein the sense strand comprises a 5’ end comprising a hydrophobic moiety, and a 3’ end comprising 2 phosphorothioate linkages; wherein any one of the following is true with regard to the sense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines, all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise 2’-O-methyl modified pyrimidines, all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines, all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines, or all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; and wherein any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’- fluoro and 2’-O-methyl modified pyrimidines, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines, all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines, all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines, or all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise 2’- fluoro modified purines.

[0266] In some embodiments, any one of the following is true with regard to the sense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise 2’-O-methyl modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; with the proviso that in any of the foregoing, the sense strand may include a deoxy nucleoside. In some embodiments, all purines comprise 2 ’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2 ’-fluoro modified purines, and all pyrimidines comprise 2’ -O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O- methyl modified purines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; with the proviso that a deoxy nucleoside may be included in the sense strand. In some embodiments, the sense strand includes the deoxy nucleoside. The deoxy nucleoside may be at nucleoside position 9 of the sense strand. In some embodiments, the sense strand does not include a deoxy nucleoside. The deoxy nucleoside of the sense strand may be otherwise unmodified. [0267] In some embodiments, any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise 2’- fluoro modified purines; with the proviso that in any of the foregoing, the sense strand may include a deoxy nucleoside. In some embodiments, all purines comprise 2 ’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise 2’-fluoro modified purines; with the proviso that a deoxy nucleoside may be included in the antisense strand. In some embodiments, the antisense strand includes the deoxy nucleoside. The deoxy nucleoside may be at nucleoside position 9 of the antisense strand. In some embodiments, the antisense strand does not include a deoxy nucleoside. The deoxy nucleoside of the antisense strand may be otherwise unmodified.

[0268] In some embodiments, any one of the following is true with regard to the sense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise 2’ -O-methyl modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; with the proviso that in any of the foregoing, the sense strand may include a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside. In some embodiments, all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’- O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise 2’ -O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’- fluoro modified pyrimidines, and all purines comprise 2’ -O-methyl modified purines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, the sense strand includes the deoxy nucleoside. The deoxy nucleoside may be at nucleoside position 9 of the sense strand. In some embodiments, the sense strand does not include a deoxy nucleoside. The deoxy nucleoside of the sense strand may be otherwise unmodified. In some embodiments, the sense strand includes a 2’-O-(2-methoxyethyl) nucleoside. The 2’-O-(2- methoxy ethyl) nucleoside may be at nucleoside position 4 of the sense strand. The 2’-O-(2- methoxy ethyl) nucleoside may include a 2’-O-(2-methoxyethyl) thymine nucleoside. In some embodiments, the sense strand does not include a 2’-O-(2-methoxyethyl) nucleoside. The 2’-O-(2- methoxy ethyl) nucleoside of the sense strand may be otherwise unmodified.

[0269] In some embodiments, any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise 2’- fluoro modified purines; with the proviso that in any of the foregoing, the sense strand may include a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside. In some embodiments, all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2- methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’- fluoro modified pyrimidines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; with the proviso that a deoxy nucleoside or a 2’-O-(2- methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; with the proviso that a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise 2’-fluoro modified purines; with the proviso that a deoxy nucleoside or a 2’- O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, the antisense strand includes the deoxy nucleoside. The deoxy nucleoside may be at nucleoside position 9 of the antisense strand. In some embodiments, the antisense strand does not include a deoxy nucleoside. The deoxy nucleoside of the antisense strand may be otherwise unmodified. In some embodiments, the antisense strand includes a 2’-O-(2-methoxyethyl) nucleoside. The 2’-O-(2- methoxy ethyl) nucleoside may be at nucleoside position 4 of the sense strand. The 2’-O-(2- methoxy ethyl) nucleoside may include a 2’-O-(2-methoxyethyl) thymine nucleoside. In some embodiments, the antisense strand does not include a 2’-O-(2-methoxyethyl) nucleoside. The 2’-O-(2- methoxy ethyl) nucleoside of the antisense strand may be otherwise unmodified.

[0270] In some embodiments, any one of the following is true with regard to the sense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise 2’-O-methyl modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; with the proviso that in any of the foregoing, the sense strand may include a 2’-O-(2-methoxyethyl) nucleoside. In some embodiments, all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a 2'-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all purines comprise 2’ -fluoro modified purines, and all pyrimidines comprise 2’ - O-methyl modified pyrimidines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’ -O-methyl modified purines; with the proviso that a 2’- O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, in the sense strand, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise 2’-O- methyl modified purines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the sense strand. In some embodiments, the sense strand includes a 2’-O-(2-methoxyethyl) nucleoside. The 2’-O-(2-methoxyethyl) nucleoside may be at nucleoside position 4 of the sense strand. The 2’-O-(2-methoxyethyl) nucleoside may include a 2’-O-(2-methoxyethyl) thymine nucleoside. In some embodiments, the sense strand does not include a 2’-O-(2-methoxyethyl) nucleoside. The 2’ -O-(2-methoxy ethyl) nucleoside of the sense strand may be otherwise unmodified. [0271] In some embodiments, any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’-fluoro modified pyrimidines; all pyrimidines comprise 2’-fluoro modified pyrimidines, and all purines comprise a mixture of 2’ -fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; or all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise 2’- fluoro modified purines; with the proviso that in any of the foregoing, the sense strand may include a deoxy nucleoside or a 2’-O-(2-methoxyethyl) nucleoside. In some embodiments, all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all purines comprise 2’ -O-methyl modified purines, and all pyrimidines comprise 2 ’-fluoro modified pyrimidines; with the proviso that a 2’ -O-(2-methoxy ethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; with the proviso that a 2’-O-(2- methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; with the proviso that a 2’-O-(2- methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, in the antisense strand, all pyrimidines comprise 2’ -O-methyl modified pyrimidines, and all purines comprise 2’ -fluoro modified purines; with the proviso that a 2’-O-(2-methoxyethyl) nucleoside may be included in the antisense strand. In some embodiments, the antisense strand includes a 2’-O-(2- methoxy ethyl) nucleoside. The 2’-O-(2-methoxyethyl) nucleoside may be at nucleoside position 4 of the sense strand. The 2’-O-(2-methoxyethyl) nucleoside may include a 2’-O-(2-methoxyethyl) thymine nucleoside. In some embodiments, the antisense strand does not include a 2’-O-(2- methoxy ethyl) nucleoside. The 2’-O-(2-methoxyethyl) nucleoside of the antisense strand may be otherwise unmodified.

[0272] In some embodiments, any one of the following is true with regard to the sense strand, with the proviso that the sense strand may include a 2’ deoxy nucleoside: all purine nucleosides comprise 2’-fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all purine nucleosides comprise 2’-fluoro, and all pyrimidine nucleosides comprise 2’-O-methyl, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O-methyl, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O- methyl, or all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides comprise 2’-O- methyl. In some embodiments, in the sense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, in the sense strand, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, in the sense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides comprise 2’-O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides comprise 2’ -O-methyl, with the proviso that the sense strand may include a 2’ deoxy nucleoside. In some embodiments, the sense strand includes the 2’ deoxy nucleoside. In some embodiments, the sense strand does not include the 2’ deoxy nucleoside. Some embodiments include a proviso that the sense strand may include a 2’-O-(2-methoxyethyl) nucleoside (e.g., at position 4, counting from 5’ to 3’). Some embodiments include the 2’-O-(2- methoxy ethyl) nucleoside in the sense strand. Some embodiments do not include the 2’-O-(2- methoxy ethyl) nucleoside in the sense strand.

[0273] In some embodiments, any one of the following is true with regard to the sense strand: all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides comprise 2’ -O-methyl, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, or all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides comprise 2’ -O-methyl. In some embodiments, in the sense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methyl. In some embodiments, in the sense strand, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl. In some embodiments, in the sense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides comprise 2’-O-methyl. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O- methyl. In some embodiments, in the sense strand, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides comprise 2’ -O-methyl. Some embodiments include a proviso that the sense strand may include a 2’-O-methoxyethyl nucleoside (e.g, at position 4, counting from 5’ to 3’). Some embodiments include the 2’-O-methoxyethyl nucleoside in the sense strand. Some embodiments do not include the 2’-O-methoxyethyl nucleoside in the sense strand.

[0274] In some embodiments, any one of the following is true with regard to the antisense strand, with the proviso that the antisense strand may include a 2’ deoxy nucleoside: all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides comprise 2’-fluoro, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methyl, or all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides comprise 2’ -fluoro. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides comprise 2’ -O-methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides comprise 2’ -O-methyl, with the proviso that the antisense strand may include a 2’ deoxy nucleoside. In some embodiments, the antisense strand includes the 2’ deoxy nucleoside. In some embodiments, the antisense strand does not include the 2’ deoxy nucleoside. Some embodiments include a proviso that the antisense strand may include a 2’ -O-methoxy ethyl nucleoside (e.g., at position 4, counting from 5’ to 3’). Some embodiments include the 2’ -O-methoxyethyl nucleoside in the antisense strand. Some embodiments do not include the 2’ -O-methoxyethyl nucleoside in the antisense strand.

[0275] In some embodiments, any one of the following is true with regard to the antisense strand: all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O-methyl, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides comprise 2’ -fluoro, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl, all pyrimidine nucleosides comprise 2’-O-methyl, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl, or all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides comprise 2’ -fluoro. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methyl. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’ -O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl. In some embodiments, in the antisense strand, all purine nucleosides comprise 2’-fluoro, and all pyrimidine nucleosides comprise 2’ -O-methyl. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides are modified with a mixture of 2’ -fluoro and 2’ -O-methyl. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -O-methyl, and all purine nucleosides are modified with a mixture of 2’- fluoro and 2’ -O-methyl. In some embodiments, in the antisense strand, all pyrimidine nucleosides comprise 2’ -fluoro, and all purine nucleosides comprise 2’ -O-methyl. Some embodiments include a proviso that the antisense strand may include a 2’ -O-methoxy ethyl nucleoside (e.g., at position 4, counting from 5’ to 3’). Some embodiments include the 2’ -O-methoxy ethyl nucleoside in the antisense strand. Some embodiments do not include the 2’ -O-methoxy ethyl nucleoside in the antisense strand.

[0276] In some embodiments, the antisense strand comprises one or two 3’ phosphorothioate linkages. For example, there may be a phosphorothioate linkage between the first and second nucleotides from the 3’ end of the antisense strand, or there may be phosphorothioate linkages between the first, second and third nucleotides from the 3’ end of the antisense strand. In some embodiments, the sense strand comprises one or two 5’ phosphorothioate linkages. For example, there may be a phosphorothioate linkage between the first and second nucleotides from the 5’ end of the sense strand, or there may be phosphorothioate linkages between the first, second and third nucleotides from the 5’ end of the sense strand. In some embodiments, the sense strand does not comprise one or two 5’ phosphorothioate linkages. For example, in some embodiments, there are no phosphorothioate linkages between the last 3 nucleotides at the 5’ end of the sense strand. In some embodiments, the sense strand comprises 5’ phosphate linkages. In some embodiments, the sense strand comprises one or two 3’ phosphorothioate linkages. For example, there may be a phosphorothioate linkage between the first and second nucleotides from the 3’ end of the sense strand, or there may be phosphorothioate linkages between the first, second and third nucleotides from the 3’ end of the sense strand.

[0277] In some embodiments, the antisense strand comprises a 5’ end comprising 2 phosphorothioate linkages. The 5’ end may comprise 3 nucleosides separated by the 2 phosphorothioate linkages. In some embodiments, the antisense strand comprises a 3’ end comprising 2 phosphorothioate linkages. The 3’ end may comprise 3 nucleosides separated by the 2 phosphorothioate linkages.

[0278] In some cases, position 9 of the sense strand can be a 2’deoxy. In these cases, 2’F and 2’0Me modifications may occur at the other positions of the sense strand. In some cases, the sense strand of any of the siRNAs comprises a modification pattern which conforms to these sense strand rules.

[0279] In some cases, the sense strand of any of the siRNAs comprises a modification pattern which conforms to these sense strand rules.

[0280] In some embodiments, the sense strand comprises or consists of RNA or modified RNA nucleotides. In some embodiments, the sense strand comprises a deoxy nucleoside. The deoxy nucleoside may include a DNA nucleoside. In some embodiments, the deoxy nucleoside comprises or consists of a 2’ deoxy nucleoside. The deoxy nucleoside may be at a position within the sense strand (5’ to 3’, where the 5’ position is 1). The position within the sense strand may be or include position 2, 4, 6, 8, 9, 10, 12, 14, 16, or 18, or a combination of said positions. The position within the sense strand may be or include position 2, 4, 6, 8, 10, 12, 14, 16, or 18, or a combination of said positions. The position within the sense strand may be or include position 2, 6, 9, 10, 14, or 18, or a combination of said positions. The position within the sense strand may be or include position 2, 6, 10, 14, or 18, or a combination of said positions. The position within the sense strand may be or include position 4, 8, 9, 12, or 16, or a combination of said positions. The position within the sense strand may be or include position 4, 8, 12, or 16, or a combination of said positions. The position within the sense strand may include position 9. The position within the sense strand may be position 9. The sense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides. In some embodiments, the sense strand includes 1 deoxy nucleoside. The sense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides, or a range of deoxy nucleosides defined by any two of the aforementioned numbers of deoxy nucleosides. The sense strand may include deoxy nucleosides at all even positions. The sense strand may include deoxy nucleosides at some even positions. The sense strand may include deoxy nucleosides at every other even position. The sense strand may include 1 deoxy nucleoside. The sense strand may include at least 1 deoxy nucleoside. The sense strand may include at least 2 deoxy nucleosides. The sense strand may include at least 3 deoxy nucleosides. The sense strand may include at least 4 deoxy nucleosides. The sense strand may include at least 5 deoxy nucleosides. The sense strand may include at least 6 deoxy nucleosides. The sense strand may include at least 7 deoxy nucleosides. The sense strand may include at least 8 deoxy nucleosides. The sense strand may include at least 9 deoxy nucleosides. The sense strand may include at least 10 deoxy nucleosides. The sense strand may include no greater than 2 deoxy nucleosides. The sense strand may include no greater than 3 deoxy nucleosides. The sense strand may include no greater than 4 deoxy nucleosides. The sense strand may include no greater than 5 deoxy nucleosides. The sense strand may include no greater than 6 deoxy nucleosides. The sense strand may include no greater than 7 deoxy nucleosides. The sense strand may include no greater than 8 deoxy nucleosides. The sense strand may include no greater than 9 deoxy nucleosides. The sense strand may include no greater than 10 deoxy nucleosides.

[0281] In some embodiments, the antisense strand comprises or consists of RNA or modified RNA nucleotides. In some embodiments, the antisense strand comprises a deoxy nucleoside. The deoxy nucleoside may include a DNA nucleoside. In some embodiments, the deoxy nucleoside comprises or consists of a2’ deoxy nucleoside. The antisense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides, or a range of deoxy nucleosides defined by any two of the aforementioned numbers of deoxy nucleosides.

[0282] In some embodiments in which a deoxy nucleoside is included in the sense strand (e.g. , at the 9th nucleotide counting from 5’ end), nucleosides at positions 1-8 include a mixture of 2’-Fluoro and 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, purines at positions 1-8 include a mixture of 2’ -fluoro and 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, pyrimidines at positions 1-8 include a mixture of 2’ -fluoro and 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, nucleosides at positions 1-8 all include 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, purines at positions 1 -8 all include 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, pyrimidines at positions 1-8 all include 2’-O-methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, purines at positions 1 -8 include a mixture of 2’-fluoro and 2’-O-methyl modified nucleosides, and pyrimidines at positions 1-8 all include 2’-O- methyl modified nucleosides. In some embodiments in which a deoxy nucleoside is included in the sense strand, pyrimidines at positions 1-8 include a mixture of 2’ -fluoro and 2’-O-methyl modified nucleosides, and purines at positions 1-8 all include 2’-O-methyl modified nucleosides.

[0283] Disclosed herein, in some embodiments are compositions comprising an oligonucleotide that targets GPAM and when administered to a cell decreases expression of GPAM, wherein the oligonucleotide comprises a small interfering RNA (siRNA) comprising a sense strand and an antisense strand, wherein the sense strand comprises a sense strand sequence described herein in which at least one internucleoside linkage is modified and at least one nucleoside is modified, or an sense strand sequence comprising 1 or 2 nucleoside substitutions, additions, or deletions of the oligonucleotide sequence in which at least one internucleoside linkage is modified and at least one nucleoside is modified, and wherein the antisense strand comprises an antisense strand sequence described herein in which at least one intemucleoside linkage is modified and at least one nucleoside is modified, or an oligonucleotide sequence comprising 1 or 2 nucleoside substitutions, additions, or deletions of the antisense strand sequence in which at least one intemucleoside linkage is modified and at least one nucleoside is modified. Some embodiments relate to methods that include administering the composition to a subject.

[0284] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 12709-12733. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 12709-12733, at least 80% identical to any one of SEQ ID NOs: 12709-12733, at least 85% identical to of any one of SEQ ID NOs: 12709-12733, at least 90% identical to any one of SEQ ID NOs: 12709-12733, or at least 95% identical to any one of SEQ ID NOs: 12709-12733. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12709-12733, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12709-12733, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 12709-12733.

[0285] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 12759-12866. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 12759-12866, at least 80% identical to any one of SEQ ID NOs: 12759-12866, at least 85% identical to of any one of SEQ ID NOs: 12759-12866, at least 90% identical to any one of SEQ ID NOs: 12759-12866, or at least 95% identical to any one of SEQ ID NOs: 12759-12866. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12759-12866, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12759-12866, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 12759-12866. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 2856-3037.

[0286] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID Nos: 12734-12758. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 12734- 12758, at least 80% identical to any one of SEQ ID Nos: 12734-12758, at least 85% identical to of any one of SEQ ID Nos: 12734-12758, at least 90% identical to any one of SEQ ID Nos: 12734- 12758, or at least 95% identical to any one of SEQ ID Nos: 12734-12758. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 12734- 12758, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 12734-12758, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 12734-12758. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 12734- 12758.

[0287] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID Nos: 12868-12974. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 12868-12974, at least 80% identical to any one of SEQ ID Nos: 12868-12974, at least 85% identical to of any one of SEQ ID Nos: 12868-12974, at least 90% identical to any one of SEQ ID Nos: 12868-12974, or at least 95% identical to any one of SEQ ID Nos: 12868-12974. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 12868-12974, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 12868-12974, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 12868-12974. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 12868-12974.

[0288] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 12975-13081. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 12975- 13081, at least 80% identical to any one of SEQ ID NOs: 12975-13081, at least 85% identical to of any one of SEQ ID NOs: 12975-13081, at least 90% identical to any one of SEQ ID NOs: 12975- 13081, or at least 95% identical to any one of SEQ ID NOs: 12975-13081. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12975- 13081, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12975-13081, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 12975-13081. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 12975- 13081.

[0289] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 13724-13751. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13724-13751, at least 80% identical to any one of SEQ ID NOs: 13724-13751, at least 85% identical to of any one of SEQ ID NOs: 13724-13751, at least 90% identical to any one of SEQ ID NOs: 13724-13751, or at least 95% identical to any one of SEQ ID NOs: 13724-13751. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13724-13751, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13724-13751, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13724-13751. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13724-13751.

[0290] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13752-13779. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13752- 13779, at least 80% identical to any one of SEQ ID NOs: 13752-13779, at least 85% identical to of any one of SEQ ID NOs: 13752-13779, at least 90% identical to any one of SEQ ID NOs: 13752- 13779, or at least 95% identical to any one of SEQ ID NOs: 13752-13779. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13752- 13779, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13752-13779, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13752-13779. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13752- 13779.

[0291] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 13780-13840. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13780-13840, at least 80% identical to any one of SEQ ID NOs: 13780-13840, at least 85% identical to of any one of SEQ ID NOs: 13780-13840, at least 90% identical to any one of SEQ ID NOs: 13780-13840, or at least 95% identical to any one of SEQ ID NOs: 13780-13840. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13780-13840, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13780-13840, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13780-13840. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13780-13840.

[0292] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13841-13913. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13841- 13913, at least 80% identical to any one of SEQ ID NOs: 13841-13913, at least 85% identical to of any one of SEQ ID NOs: 13841-13913, at least 90% identical to any one of SEQ ID NOs: 13841- 13913, or at least 95% identical to any one of SEQ ID NOs: 13841-13913. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13841- 13913, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13841-13913, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13841-13913. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13841- 13913.

[0293] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID Nos: 13914-13937. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 13914-13937, at least 80% identical to any one of SEQ ID Nos: 13914-13937, at least 85% identical to of any one of SEQ ID Nos: 13914-13937, at least 90% identical to any one of SEQ ID Nos: 13914-13937, or at least 95% identical to any one of SEQ ID Nos: 13914-13937. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 13914-13937, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 13914-13937, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 13914-13937. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13914-13937.

[0294] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13938-13950. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13938- 13950, at least 80% identical to any one of SEQ ID NOs: 13938-13950, at least 85% identical to of any one of SEQ ID NOs: 13938-13950, at least 90% identical to any one of SEQ ID NOs: 13938- 13950, or at least 95% identical to any one of SEQ ID NOs: 13938-13950. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13938- 13950, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13938-13950, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13938-13950. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 13938- 13950.

[0295] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333, at least 80% identical to any one of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333, at least 85% identical to of any one of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333, at least 90% identical to any one of SEQ ID Nos:

14254-14255, 14308-14319, or 14331-14333, or at least 95% identical to any one of SEQ ID Nos:

14254-14255, 14308-14319, or 14331-14333. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 14254-14255, 14308-14319, or 14331-14333. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep- 2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 14254-14255, 14308-14319, or 14331-14333.

[0296] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 14207-14253, 14256-14284, 14320-14330, or 14334-14336. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 14207-14253, 14256-14284, 14320-14330, or 14334-14336, at least 80% identical to any one of SEQ ID NOs: 14207-14253, 14256-14284, 14320-14330, or 14334- 14336, at least 85% identical to of any one of SEQ ID NOs: 14207-14253, 14256-14284, 14320- 14330, or 14334-14336, at least 90% identical to any one of SEQ ID NOs: 14207-14253, 14256- 14284, 14320-14330, or 14334-14336, or at least 95% identical to any one of SEQ ID NOs: 14207- 14253, 14256-14284, 14320-14330, or 14334-14336. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14207-14253, 14256- 14284, 14320-14330, or 14334-14336, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 14207-14253, 14256-14284, 14320-14330, or 14334-14336, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 14207-14253, 14256-14284, 14320-14330, or 14334-14336. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein, such as a different set of modifications or modification pattern than the aforementioned sequences. The antisense strand may comprise a lipid moiety or a GalNAc moiety or a lipid moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to antisense strand sequence of one of SEQ ID NO: 14207-14253, 14256-14284, 14320-14330, or 14334-14336.

[0297] In some embodiments, any of the aforementioned siRNA comprising a sense stand comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NOS: 12709-12733, 12868-12974, 13724-13751, 13724- 13751, 13780-13840, 13914-13937, 14254-14255, 14308-14319 or 14331-14333. The sense strand may comprise a lipid moiety. The sense strand may comprise a GalNAc moiety. The sense strand may comprise an integrin or an integrin targeting ligand. The sense strand may comprise an angiopep-2. The sense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The sense strand may comprise a glucose transporter protein. The sense strand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6] s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28. Preferably, the lipid moiety is ETL20.

Representative examples of integrin or integrin targeting ligand is epithelial -specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha- v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0298] In some embodiments, any of the aforementioned siRNA comprising an antisense stand comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand sequence of one of SEQ ID NOs: 12734-12758, 12975-13081, 13752-13723, 13752- 13779, 13841-13913, 13938-13950, 14207-14253, 14256-14284, 14320-14330 or 14334-14336. The antisense stand may comprise a lipid moiety. The antisense strand may comprise a GalNAc moiety. The antisense strand may comprise an integrin or an integrin targeting ligand. The antisense strand may comprise an angiopep-2. The antisense strand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The antisense strand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23, L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6] s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28.

[0299] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset G. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset G. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset G, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset G, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset G. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset G). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset G. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of subset G. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of subset G.

[0300] In some embodiments, the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset H. In some embodiments, the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset H. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset H, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset H, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset H. The sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences. The sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand or antisense strand may comprise an overhang. The sense strand or antisense strand may comprise any modifications described herein (e.g., a different set of modifications or modification pattern than subset H). The sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense strand or antisense strand sequence of subset H. Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of subset H. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5’ U of an antisense strand sequence of subset H.

[0301] In some embodiments, any of the aforementioned siRNA comprising a sense strand or antisense stand comprises a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand attached to sense or anti sense strand sequence of one of Subset G, or subset H. The sense strand or antisense stand may comprise a lipid moiety. The antisense strand may comprise a GalNAc moiety, sense strand or antisense stand may comprise an integrin or an integrin targeting ligand. The sense strand or antisense stand may comprise an angiopep-2. The sense strand or antisense stand may comprise a lipoprotein receptor related protein (LRP) ligand. The antisense strand may comprise a glucose transporter protein. The sense strand or antisense stand may comprise a LDL receptor ligand. Representative examples of the GalNAc moiety includes, but is not limited to, ETL1, ETL17, NAG37, ST23, GluGalNAc, K2GalNAc, PyrGalNAc, PipGalNAc, TEG-GalNAc, GalNAc23 , L-9, Sirius GalNAc, GLS-5, GLS-15, Olix GalNAc, lgT3, 5gn2c6, [Gal-6] s[Gal-6]s[Gal-6], Janssen, Arbutus or THA. Preferably, the GalNAc moiety is ETL17. Representative examples of lipid moiety includes, but is not limited to, ETL3, ETL7, ETL8, ETL9, ETL10, ETL12, ETL13, ETL15, ETL16, ETL18, ETL19, ETL20, ETL21, ETL22 or ETL28. Preferably, the lipid moiety is ETL20. Representative examples of integrin or integrin targeting ligand is epithelial-specific integrin, integrin alpha-v-beta-6 (αvβ6) or integrin alpha- v-beta-3 or arginine-glycine-aspartic acid (RGD) peptide. In any of SEQ ID NOs: 13951-14078, thymine (T) may be replaced with uracil (U).

[0302] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735, 12959, 13736, 13737, 13738, 13739, 13740, 13741, 13742, 13743,

13744, 13745, 13746, 13747, 13815, 13972, 13822, 12877, 12927, 12928, 12929, 12930, 12931,

12932, 12933, 12934, 12960, 13748, 13749, 13750, 13751, 12883, 12910, 12911, 12912, 12913,

12914, 12915, 12916, 12917, 12947, 12948, 12965, 12885, 12901, 12902, 12903, 12904, 12905,

12906, 12907, 12908, 12909, 12886, 12893, 12894, 12895, 12896, 12897, 12898, 12899, 12900,

12968, 12889, 12918, 12919, 12920, 12921, 12922, 12923, 12924, 12925, 12926, 12943, 12944,

12945, 12946, 12969, 12970, 12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735, 12959, 13736, 13737, 13738, 13739, 13740, 13741,

13742, 13743, 13744, 13745, 13746, 13747, 13815, 13972, 13822, 12877, 12927, 12928, 12929,

12930, 12931, 12932, 12933, 12934, 12960, 13748, 13749, 13750, 13751, 12883, 12910, 12911,

12912, 12913, 12914, 12915, 12916, 12917, 12947, 12948, 12965, 12885, 12901, 12902, 12903,

12904, 12905, 12906, 12907, 12908, 12909, 12886, 12893, 12894, 12895, 12896, 12897, 12898,

12899, 12900, 12968, 12889, 12918, 12919, 12920, 12921, 12922, 12923, 12924, 12925, 12926,

12943, 12944, 12945, 12946, 12969, 12970, 12892, 12935, 12936, 12937, 12938, 12939, 12940,

12941, or 12942, at least 80% identical to any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735, 12959, 13736, 13737,

13738, 13739, 13740, 13741, 13742, 13743, 13744, 13745, 13746, 13747, 13815, 13972, 13822,

12877, 12927, 12928, 12929, 12930, 12931, 12932, 12933, 12934, 12960, 13748, 13749, 13750,

13751, 12883, 12910, 12911, 12912, 12913, 12914, 12915, 12916, 12917, 12947, 12948, 12965,

12885, 12901, 12902, 12903, 12904, 12905, 12906, 12907, 12908, 12909, 12886, 12893, 12894,

12895, 12896, 12897, 12898, 12899, 12900, 12968, 12889, 12918, 12919, 12920, 12921, 12922,

12923, 12924, 12925, 12926, 12943, 12944, 12945, 12946, 12969, 12970, 12892, 12935, 12936,

12937, 12938, 12939, 12940, 12941, or 12942, at least 85% identical to of any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735, 12959, 13736, 13737, 13738, 13739, 13740, 13741, 13742, 13743, 13744, 13745, 13746,

13747, 13815, 13972, 13822, 12877, 12927, 12928, 12929, 12930, 12931, 12932, 12933, 12934,

12960, 13748, 13749, 13750, 13751, 12883, 12910, 12911, 12912, 12913, 12914, 12915, 12916,

12917, 12947, 12948, 12965, 12885, 12901, 12902, 12903, 12904, 12905, 12906, 12907, 12908,

12909, 12886, 12893, 12894, 12895, 12896, 12897, 12898, 12899, 12900, 12968, 12889, 12918,

12919, 12920, 12921, 12922, 12923, 12924, 12925, 12926, 12943, 12944, 12945, 12946, 12969,

12970, 12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942, at least 90% identical to any one of SEQ ID NOs: 13403-13723, or at least 95% identical to any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735,

12959, 13736, 13737, 13738, 13739, 13740, 13741, 13742, 13743, 13744, 13745, 13746, 13747,

13815, 13972, 13822, 12877, 12927, 12928, 12929, 12930, 12931, 12932, 12933, 12934, 12960,

13748, 13749, 13750, 13751, 12883, 12910, 12911, 12912, 12913, 12914, 12915, 12916, 12917,

12947, 12948, 12965, 12885, 12901, 12902, 12903, 12904, 12905, 12906, 12907, 12908, 12909,

12886, 12893, 12894, 12895, 12896, 12897, 12898, 12899, 12900, 12968, 12889, 12918, 12919,

12920, 12921, 12922, 12923, 12924, 12925, 12926, 12943, 12944, 12945, 12946, 12969, 12970,

12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735,

12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735,

12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 12953, 12958, 13724, 13725, 13726, 13727, 13728, 13729, 13730, 13731, 13732, 13733, 13734, 13735,

12892, 12935, 12936, 12937, 12938, 12939, 12940, 12941, or 12942. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein. The sense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, or LDL receptor ligand.

[0303] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984, 13034, 13035, 13036, 13037,

13038, 13039, 13040, 13041, 13067, 13765, 13766, 13767, 13768, 12990, 13017, 13018, 13019,

13020, 13021, 13022, 13023, 13024, 13054, 13055, 13072, 12992, 13008, 13009, 13010, 13011,

13012, 13013, 13014, 13015, 13016, 12993, 13000, 13001, 13002, 13003, 13004, 13005, 13006,

13007, 13075, 12996, 13025, 13026, 13027, 13028, 13029, 13030, 13031, 13032, 13033, 13050,

13051, 13052, 13053, 13076, 13077, 12999, 13042, 13043, 13044, 13045, 13046, 13047, 13048,

13049, 14219, 14277, 14229, or 14284. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066,

13764, 12984, 13034, 13035, 13036, 13037, 13038, 13039, 13040, 13041, 13067, 13765, 13766,

13767, 13768, 12990, 13017, 13018, 13019, 13020, 13021, 13022, 13023, 13024, 13054, 13055,

13072, 12992, 13008, 13009, 13010, 13011, 13012, 13013, 13014, 13015, 13016, 12993, 13000,

13001, 13002, 13003, 13004, 13005, 13006, 13007, 13075, 12996, 13025, 13026, 13027, 13028,

13029, 13030, 13031, 13032, 13033, 13050, 13051, 13052, 13053, 13076, 13077, 12999, 13042,

13043, 13044, 13045, 13046, 13047, 13048, 13049, 14219, 14277, 14229, or 14284, at least 80% identical to any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757,

13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984, 13034, 13035, 13036, 13037,

13020, 13021, 13022, 13023, 13024, 13054, 13055, 13072, 12992, 13008, 13009, 13010, 13011, 13012, 13013, 13014, 13015, 13016, 12993, 13000, 13001, 13002, 13003, 13004, 13005, 13006,

13049, 14219, 14277, 14229, or 14284, at least 85% identical to of any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763,

13066, 13764, 12984, 13034, 13035, 13036, 13037, 13038, 13039, 13040, 13041, 13067, 13765,

13766, 13767, 13768, 12990, 13017, 13018, 13019, 13020, 13021, 13022, 13023, 13024, 13054,

13055, 13072, 12992, 13008, 13009, 13010, 13011, 13012, 13013, 13014, 13015, 13016, 12993,

13000, 13001, 13002, 13003, 13004, 13005, 13006, 13007, 13075, 12996, 13025, 13026, 13027,

13028, 13029, 13030, 13031, 13032, 13033, 13050, 13051, 13052, 13053, 13076, 13077, 12999,

13042, 13043, 13044, 13045, 13046, 13047, 13048, 13049, 14219, 14277, 14229, or 14284, at least 90% identical to any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984, 13034, 13035, 13036, 13037,

13049, 14219, 14277, 14229, or 14284, or at least 95% identical to any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763,

13042, 13043, 13044, 13045, 13046, 13047, 13048, 13049, 14219, 14277, 14229, or 14284. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984, 13034, 13035, 13036, 13037, 13038, 13039, 13040, 13041,

13067, 13765, 13766, 13767, 13768, 12990, 13017, 13018, 13019, 13020, 13021, 13022, 13023,

13024, 13054, 13055, 13072, 12992, 13008, 13009, 13010, 13011, 13012, 13013, 13014, 13015,

13016, 12993, 13000, 13001, 13002, 13003, 13004, 13005, 13006, 13007, 13075, 12996, 13025,

13026, 13027, 13028, 13029, 13030, 13031, 13032, 13033, 13050, 13051, 13052, 13053, 13076,

13077, 12999, 13042, 13043, 13044, 13045, 13046, 13047, 13048, 13049, 14219, 14277, 14229, or 14284, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984, 13034, 13035, 13036, 13037,

13049, 14219, 14277, 14229, or 14284, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13060, 13065, 13752, 13753, 13754, 13755, 13756, 13757, 13758, 13759, 13760, 13761, 13762, 13763, 13066, 13764, 12984,

13034, 13035, 13036, 13037, 13038, 13039, 13040, 13041, 13067, 13765, 13766, 13767, 13768,

12990, 13017, 13018, 13019, 13020, 13021, 13022, 13023, 13024, 13054, 13055, 13072, 12992,

13008, 13009, 13010, 13011, 13012, 13013, 13014, 13015, 13016, 12993, 13000, 13001, 13002,

13003, 13004, 13005, 13006, 13007, 13075, 12996, 13025, 13026, 13027, 13028, 13029, 13030,

13031, 13032, 13033, 13050, 13051, 13052, 13053, 13076, 13077, 12999, 13042, 13043, 13044,

13045, 13046, 13047, 13048, 13049, 14219, 14277, 14229, or 14284. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, or LDL receptor ligand.

[0304] In some embodiments, the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822. In some embodiments, the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822, at least 80% identical to any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822, at least 85% identical to of any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822, at least 90% identical to any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822,, or at least 95% identical to any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 12959, 12928, 12932, 12960, 12947, 12965, 12904, 12909, 12897, 12968, 12926, 12969, 12970, 12935, 12936, 13815, 13972, or 13822. The sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The sense strand may comprise an overhang. The sense strand may comprise a modification pattern described herein. The sense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, or LDL receptor ligand.

[0305] In some embodiments, the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284. In some embodiments, the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, at least 80% identical to any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, at least 85% identical to of any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, at least 90% identical to any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, or at least 95% identical to any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 13060, 13065, 13066, 13035, 13039, 13067, 13054, 13072, 13011, 13016, 13004, 13075, 13033, 13076, 13077, 13042, 13043, 14219, 14229, 14277, or 14284. The antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences. The antisense strand may comprise an overhang. The antisense strand may comprise a modification pattern described herein. The antisense strand may comprise a lipid moiety or a GalNAc moiety or integrin or integrin targeting ligand or angiopep-2, lipoprotein receptor related protein (LRP) ligand, bEnd.3 cell binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, or LDL receptor ligand.

4. Modified ASOs

In some embodiments, the composition comprises an oligonucleotide that inhibits the expression of GPAM, wherein the oligonucleotide comprises an antisense oligonucleotide (ASO). In some embodiments, the ASO comprises modification pattern ASO1:

(SEQ ID NO: 12928), wherein “dN” is any deoxynucleotide, “n” is a 2’-O-methyl or 2’-O-(2-methoxyethyl)-modified nucleoside, and “s” is a phosphorothioate linkage. In some embodiments, the ASO comprises modification pattan 1S, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 11S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 1AS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, 13AS, 14AS, 15AS, 16AS, 17AS, 18AS, 19AS, 20AS, 21 AS, 22AS, 23 AS, 24AS, 25AS, 26AS, 27 AS, 28AS, 29AS, 30AS, 3 IAS, 32AS, 33AS, 34AS, 35AS, 36AS, 37AS, 38AS, 39AS, or 40AS.

D. F ormulations

[0306] In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition is sterile. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0307] In some embodiments, the pharmaceutically acceptable carrier comprises water. In some embodiments, the pharmaceutically acceptable carrier comprises a buffer. In some embodiments, the pharmaceutically acceptable carrier comprises a saline solution. In some embodiments, the pharmaceutically acceptable carrier comprises water, a buffer, or a saline solution. In some embodiments, the composition comprises a liposome. In some embodiments, the pharmaceutically acceptable carrier comprises liposomes, lipids, nanoparticles, proteins, protein-antibody complexes, peptides, cellulose, nanogel, or a combination thereof.

E. Kits

[0308] Described herein, in some embodiments, are kits. The kit may include an oligonucleotide such as an siRNA described herein. The oligonucleotide may be conjugated to a lipid moiety or to a sugar moiety. The kit may include a lipid moiety. The kit may include a sugar moiety. The oligonucleotide may comprise nucleoside modifications or modified internucleoside linkages. The oligonucleotide may include any modifications described herein, such as modifications from a base sequence. The kit may include a delivery reagent such as a needle. The kit may include instructions for use, such as methods for use in a method described herein.

II. METHODS AND USES

[0309] Disclosed herein, in some embodiments, are methods of administering a composition described herein to a subject. Some embodiments relate to use a composition described herein, such as administering the composition to a subject.

[0310] Some embodiments relate to a method of treating a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of treatment. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration treats the disorder in the subject. In some embodiments, the composition treats the disorder in the subject. The disorder may comprise a disease.

[0311] In some embodiments, the treatment comprises prevention, inhibition, improvement, or reversion of the disorder in the subject. Some embodiments relate to use of a composition described herein in the method of preventing, inhibiting, improving, or reversing the disorder. Some embodiments relate to a method of preventing, inhibiting, improving, or reversing a disorder a disorder in a subject in need thereof. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration prevents, inhibits, improves, or reverses the disorder in the subject. In some embodiments, the composition prevents, inhibits, improves, or reverses the disorder in the subject.

[0312] Some embodiments relate to a method of preventing a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of preventing the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration prevents the disorder in the subject. In some embodiments, the composition prevents the disorder in the subject.

[0313] Some embodiments relate to a method of inhibiting a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of inhibiting the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration inhibits the disorder in the subject. In some embodiments, the composition inhibits the disorder in the subject.

[0314] Some embodiments relate to a method of reversing a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of reversing the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration reverses the disorder in the subject. In some embodiments, the composition reverses the disorder in the subject.

[0315] Some embodiments relate to a method of improving a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of improving the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration improves the disorder in the subject. In some embodiments, the composition improves the disorder in the subject.

[0316] Some embodiments relate to a method of treating a subject having liver disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one additional active agent. Some embodiments relate to a method of treating a subject having liver disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one GLP-1 receptor agonist. In some embodiments, the liver disease comprises NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, tirzepatide, dulaglutide, semaglutide, danuglipron, orforflipron, or a combination thereof.

[0317] Some embodiments relate to a method of treating a subject having liver disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one GLP-1 receptor agonist, glucosedependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor- 15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator- activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activinE expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxy steroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof. In some embodiments, the liver disease comprises NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

[0318] Some embodiments relate to a method of treating a subject having cardiometabolic disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one additional active agent. Some embodiments relate to a method of treating subj ect having cardiometabolic disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one GLP-1 receptor agonist. In some embodiments, the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease. In some embodiments, the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, , tirzepatide, dulaglutide, semaglutide, danuglipron, orforflipron, or a combination thereof.

[0319] Some embodiments relate to a method of treating subject having cardiometabolic disease, comprising administering an effective amount of a composition described herein to the subject either alone or in combination with a therapeutically effective amount of at least one GLP-1 receptor agonist, glucose- dependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor- 15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator-activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activin E expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof. In some embodiments, the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease. [0320] In some embodiments, the composition of the present invention, can be widely combined with other pharmacologically active compounds for e.g., dual agonists for the GLP-1 and GIP receptors, GLP-1 receptor agonists and GIP receptor antagonists, or trigonal agonists for the GLP-1, GIP and glucagon receptors, and other pharmacologically active compounds e.g., amylin receptor agonists, apelin receptor agonists, peptide YY receptor agonists. In some embodiments, the combinations can be used especially for an additive or synergistic improvement in action. They can be applied either by separate administration of the active ingredients to the patient or in the form of combination products in which a plurality of active ingredients are present in one pharmaceutical preparation. In some embodiments, when the active ingredients are administered by separate administration of the active ingredients, this can be done simultaneously or successively.

[0321] In some embodiments, the term “GLP-1 receptor agonist” includes GLP-1, analogs and derivatives thereof, exendin-3 and analogs and derivatives thereof, and exendin-4 and analogs and derivatives thereof. The compositions of the invention comprise one or more selected independently of one another from the group consisting of glucagon-like peptide-1 (GLP-1), analogs and derivatives of GLP-1, exendin-3, analogs and derivatives of exendin-3, exendin-4, analogs and derivatives of exendin-4, and pharmacologically tolerable salts thereof. Also included are substances which exhibit the biological activity of GLP-1.

[0322] In some embodiments, the administration is systemic. In some embodiments, the administration is by parenteral administration. In some embodiments, the administration is local (e.g., to a particular organ or tissue). In some embodiments, the administration is by inhalation. In some embodiments, the administration is topical. In some embodiments, the administration is by infusion. In some embodiments, the administration is by injection. In some embodiments, the administration is intravenous (e.g., by intravenous injection or infusion). In some embodiments, the administration is subcutaneous (e.g., by subcutaneous injection). In some embodiments, the administration is intraperitoneal. In some embodiments, the administration is intraparenchymal. In some embodiments, the administration is intramuscular (e.g., by intramuscular injection). In some embodiments, the administration may be to an eye tissue (e.g., by intravitreal, intracameral or subconjunctival injection, or by topical administration to the eye). In some embodiments, the administration may be to a central nervous system tissue (e.g., brain or spinal cord or spinal canal). In some embodiments, the administration is intracerebroventricular or intrathecal (e.g., by intrathecal injection or infusion). In some embodiments, the administration may be to joint tissue (e.g., by intra-articular injection). In some embodiments, the administration may be to airway or lung tissue (e.g., by intranasal, intratracheal or inhaled administration). In some embodiments, the administration may be to skin or connective tissue (e.g., by topical administration or injection to the skin).

A. Disorders

[0323] Some embodiments of the methods described herein include treating a disorder in a subj ect in need thereof. In some embodiments, the disorder is a liver disease. Non-limiting examples of liver diseases may include non-alcoholic fatty liver disease (NAFLD, also known as metabolic dysfunction-associated steatotic liver disease [MASLD]), non-alcoholic steatohepatitis (NASH, also known as metabolic dysfunction-associated steatohepatitis [MASH]), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma. In some embodiments, the disorder is a cardiometabolic disease. Non-limiting examples of cardiometabolic diseases may include hyperlipidemia, ischemic heart disease, or coronary heart disease. In some embodiments, the disorder comprises NAFLD (or MASLD). In some embodiments, the disorder comprises NASH (or MASH). In some embodiments, the disorder comprises alcoholic liver disease. In some embodiments, the disorder comprises liver fibrosis. In some embodiments, the disorder comprises liver cirrhosis. In some embodiments, the disorder comprises hepatocellular carcinoma. In some embodiments, the disorder comprises hyperlipidemia. In some embodiments, the disorder comprises a heart disease. In some embodiments, the heart disease comprises ischemic heart disease. In some embodiments, the heart disease comprises coronary heart disease.

B. Subjects

[0324] Some embodiments of the methods described herein include treatment of a subject. Nonlimiting examples of subjects include vertebrates, animals, mammals, dogs, cats, cattle, rodents, mice, rats, primates, monkeys, and humans. In some embodiments, the subject is a vertebrate. In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal. In some embodiments, the subject is a dog. In some embodiments, the subject is a cat. In some embodiments, the subject is a cattle. In some embodiments, the subject is a mouse. In some embodiments, the subject is a rat. In some embodiments, the subject is a primate. In some embodiments, the subject is a monkey. In some embodiments, the subject is an animal, a mammal, a dog, a cat, cattle, a rodent, a mouse, a rat, a primate, or a monkey. In some embodiments, the subject is a human.

[0325] In some embodiments, the subject is male. In some embodiments, the subject is female. In some embodiments, the subject is an adult (e.g., at least 18 years old).

[0326] In some embodiments, the subject has a body mass index (BMI) of 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, or more, or a range defined by any two of the aforementioned integers. In some embodiments, the subject is overweight. In some embodiments, the subject has a BMI of 25 or more. In some embodiments, the subject has a BMI of 25-29. In some embodiments, the subject is obese. In some embodiments, the subject has a BMI of 30 or more. In some embodiments, the subject has a BMI of 30-39. In some embodiments, the subject has a BMI of 40-50. In some embodiments, the subject has a BMI of 25-50.

C. Baseline measurements

[0327] Some embodiments of the methods described herein include obtaining a baseline measurement from a subject. For example, in some embodiments, a baseline measurement is obtained from the subject prior to treating the subject. Non-limiting examples of baseline measurements include a baseline liver fat percentage measurement, a baseline liver fibrosis score measurement, a baseline NAFLD activity score measurement, a baseline blood alanine aminotransferase (ALT) measurement, a baseline blood aspartate aminotransferase (AST) measurement, a baseline blood alkaline phosphatase (ALP) measurement, a baseline blood bilirubin measurement, a baseline low- density lipoprotein (LDL) measurement, a baseline total cholesterol measurement, a baseline non- HDL cholesterol measurement, a baseline apolipoprotein B (APOB), a baseline GPAM protein measurement, or a baseline GPAM mRNA measurement.

[0328] In some embodiments, the baseline measurement is obtained directly from the subject. In some embodiments, the baseline measurement is obtained by observation, for example by observation of the subject or of the subject’s tissue. In some embodiments, the baseline measurement is obtained noninvasively using an imaging device.

[0329] In some embodiments, the baseline measurement is obtained in a sample from the subject. In some embodiments, the baseline measurement is obtained in one or more histological tissue sections. In some embodiments, the baseline measurement is obtained by performing an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay, on the sample obtained from the subject. In some embodiments, the baseline measurement is obtained by an immunoassay, a colorimetric assay, a fluorescence assay, or a chromatography (e.g., HPLC) assay. In some embodiments, the baseline measurement is obtained by PCR.

[0330] In some embodiments, the baseline measurement is a baseline liver fat percentage measurement. In some embodiments, the baseline liver fat percentage measurement is a baseline percentage as measured by MRI. In some embodiments, the baseline liver fat percentage measurement is a baseline percentage as measured by CT scan. In some embodiments, the baseline liver fat percentage is measured in a liver biopsy.

[0331] In some embodiments, the baseline measurement is a baseline NAFLD activity score measurement. In some embodiments, the baseline NAFLD activity score measurement is measured by histological analysis. In some embodiments, the baseline NAFLD activity score measurement includes a numerical score for steatosis (0-3), hepatocyte ballooning (1-2), and lobular inflammation (0-3). In some embodiments, the score thresholds of < 3 correlates with a diagnosis of not -NASH. In some embodiments, the score thresholds of > 5 correlates with a diagnosis of NASH. In some embodiments, a baseline NAFLD activity score measurement is assayed by imaging.

[0332] In some embodiments, the baseline measurement is a baseline alanine aminotransferase (ALT) measurement. In some embodiments, the baseline ALT measurement includes a baseline blood ALT measurement. In some embodiments, the baseline ALT measurement is measured by enzymatic activity. In some embodiments, the baseline ALT measurement is measured by presence of an ALT epitope. In some embodiments, the baseline ALT measurement is a baseline circulating ALT measurement. In some embodiments, the baseline ALT measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0333] In some embodiments, the baseline measurement is a baseline aspartate aminotransferase (AST) measurement. In some embodiments, the baseline AST measurement includes a baseline blood AST measurement. In some embodiments, the baseline AST measurement is measured by enzymatic activity. In some embodiments, the baseline AST measurement is measured by presence of an AST epitope. In some embodiments, the baseline AST measurement is a baseline circulating AST measurement. In some embodiments, the baseline AST measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0334] In some embodiments, the baseline measurement is a baseline alkaline phosphatase (ALP) measurement. In some embodiments, the baseline ALP measurement includes a baseline blood ALP measurement. In some embodiments, the baseline ALP measurement is measured by enzymatic activity. In some embodiments, the baseline ALP measurement is measured by presence of an ALP epitope. In some embodiments, the baseline ALP measurement is a baseline circulating ALP measurement. In some embodiments, the baseline ALP measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0335] In some embodiments, the baseline measurement is a baseline bilirubin measurement. In some embodiments, the baseline bilirubin measurement includes a baseline blood bilirubin measurement. In some embodiments, the baseline bilirubin measurement is measured by presence of a bilirubin epitope. In some embodiments, the baseline bilirubin measurement is a baseline circulating bilirubin measurement. In some embodiments, the baseline bilirubin measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0336] In some embodiments, the baseline measurement is a baseline low-density lipoprotein (LDL) measurement. In some embodiments, the baseline LDL measurement includes a baseline blood LDL measurement. In some embodiments, the baseline LDL measurement is measured by presence of an LDL epitope. In some embodiments, the baseline LDL measurement is a baseline circulating LDL measurement. In some embodiments, the baseline LDL measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0337] In some embodiments, the baseline measurement is a baseline total cholesterol measurement. In some embodiments, the baseline total cholesterol measurement includes a baseline blood total cholesterol measurement. In some embodiments, the baseline total cholesterol is measured by presence of a total cholesterol epitope. In some embodiments, the baseline total cholesterol measurement is a baseline circulating total cholesterol measurement. In some embodiments, the baseline total cholesterol measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0338] In some embodiments, the baseline measurement is a baseline non-HDL cholesterol measurement. In some embodiments, the baseline non-HDL cholesterol measurement includes a baseline blood non-HDL cholesterol measurement. In some embodiments, the baseline non-HDL cholesterol measurement is a baseline circulating non-HDL cholesterol measurement. In some embodiments, the baseline non-HDL cholesterol measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0339] In some embodiments, the baseline measurement is a baseline apolipoprotein B (APOB) measurement. In some embodiments, the baseline APOB includes a baseline blood APOB measurement. In some embodiments, the baseline APOB is measured by presence of an APOB epitope. In some embodiments, the baseline APOB measurement is a baseline circulating APOB measurement. In some embodiments, the baseline APOB measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. [0340] In some embodiments, the baseline measurement is a baseline liver fibrosis measurement. In some embodiments, the baseline liver fibrosis measurement is a baseline liver fibrosis score (LFS). In some embodiments, the baseline LFS comprises a score of 0, 1, 2, 3, or 4, or a range of scores defined by any two of the aforementioned numbers. In some embodiments, the baseline LFS comprises a score of 0-4. In some embodiments, the baseline LFS is obtained using a scoring system exemplified in Table 2. In some embodiments, the baseline LFS measurement is obtained noninvasively. In some embodiments, the baseline LFS measurement is obtained by a medical imaging device such as a vibration-controlled transient elastography (VCTE) device, a shear wave elastography device, a medical resonance imaging (MRI) device, a magnetic resonance spectroscopy device, a computed tomography device, or an ultrasound device. In some embodiments, the baseline LFS measurement is obtained in a liver sample. In some embodiments, the baseline LFS is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. In some embodiments, the baseline LFS is obtained using one or more indirect markers or measures of liver fibrosis such as an aspartate aminotransferase-to-platelet ratio index (APRI), a Fibrosis-4 (FIB-4) index, a Fibrolndex, a Foms Index, aHepascore, or aFibroTest. In some embodiments, the baseline LFS is obtained using one or more indirect markers or measures of liver fibrosis such as a FIBROSpect test or a FIBROSpect II test. In some embodiments, the baseline LFS is obtained by RT-qPCR or RNA sequencing of one or more fibrosis-related genes such as a collagen gene. In some embodiments, the baseline LFS or the baseline LFS is obtained using a scoring system upon a visual inspection of a sample such as a histological sample. In some embodiments, the baseline LFS or the baseline LFS is obtained using a stain with an affinity to collagen. In some embodiments, the baseline LFS measurement is measured by FibroScan® in kilopascals (kPa). In some embodiments, the baseline LFS is measured by histological analysis. In some embodiments, the baseline LFS measurement is a baseline LFS measurement assayed by imaging. In some embodiments, the imaging is by MRI or CT scan.

Table 2. Non-Limiting Examples of Liver Fibrosis Scoring Systems

[0341] In some embodiments, the baseline measurement is a baseline GPAM mRNA measurement. In some embodiments, the baseline GPAM mRNA measurement comprises a baseline GPAM mRNA level. In some embodiments, the baseline GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per sample weight. In some embodiments, the baseline GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per sample volume. In some embodiments, the baseline GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per total mRNA within the sample. In some embodiments, the baseline GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per total nucleic acids within the sample. In some embodiments, the baseline GPAM mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the baseline GPAM mRNA measurement is a baseline tissue GPAM mRNA measurement. In some embodiments, the baseline GPAM mRNA measurement is obtained by an assay such as a polymerase chain reaction (PCR) assay. In some embodiments, the PCR comprises quantitative PCR (qPCR). In some embodiments, the PCR comprises reverse transcription of the GPAM mRNA.

[0342] In some embodiments, the baseline measurement is a baseline GPAM protein measurement. In some embodiments, the baseline GPAM protein measurement comprises a baseline GPAM protein level. In some embodiments, the baseline GPAM protein level is indicated as a mass or percentage of GPAM protein per sample weight. In some embodiments, the baseline GPAM protein level is indicated as a mass or percentage of GPAM protein per sample volume. In some embodiments, the baseline GPAM protein level is indicated as a mass or percentage of GPAM protein per total protein within the sample. In some embodiments, the baseline GPAM protein measurement is a baseline tissue GPAM protein measurement. In some embodiments, the baseline GPAM protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. [0343] Some embodiments of the methods described herein include obtaining a sample from a subject. In some embodiments, the baseline measurement is obtained in a sample obtained from the subject. In some embodiments, the sample is obtained from the subject prior to administration or treatment of the subject with a composition described herein. In some embodiments, a baseline measurement is obtained in a sample obtained from the subject prior to administering the composition to the subject. In some embodiments, the sample is obtained from the subject in a fasted state. In some embodiments, the sample is obtained from the subject after an overnight fasting period. In some embodiments, the sample is obtained from the subject in a fed state.

[0344] In some embodiments, the sample comprises a fluid. In some embodiments, the sample is a fluid sample. In some embodiments, the sample is a blood, plasma, or serum sample. In some embodiments, the sample comprises blood. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a whole-blood sample. In some embodiments, the blood is fractionated or centrifuged. In some embodiments, the sample comprises plasma. In some embodiments, the sample is a plasma sample. A blood sample may be a plasma sample. In some embodiments, the sample comprises serum. In some embodiments, the sample is a serum sample. A blood sample may be a serum sample.

[0345] In some embodiments, the sample comprises a tissue. In some embodiments, the sample is a tissue sample. In some embodiments, the tissue comprises liver or adipose tissue. For example, the baseline GPAM mRNA measurement, or the baseline GPAM protein measurement, may be obtained in a liver or adipose sample obtained from the patient. In some embodiments, the tissue comprises adipose tissue. In some embodiments, the adipose tissue comprises white adipose tissue. The adipose tissue may include adipocytes. The adipose tissue may include preadipocytes In some embodiments, the tissue comprises liver tissue. The liver may include hepatocytes. In some embodiments, the tissue comprises vasculature tissue. In some embodiments, the vasculature tissue comprises endothelial cells.

[0346] In some embodiments, the sample includes cells. In some embodiments, the sample comprises a cell. In some embodiments, the cell comprises an adipose cell or a liver cell. In some embodiments, the cell is an adipose cell. In some embodiments, the adipose cell is an adipocyte. In some embodiments, the cell is a liver cell. In some embodiments, the liver cell is a hepatocyte. In some embodiments, the cell is a vasculature cell. In some embodiments, the vasculature cell is an endothelial cell.

D. Effects

[0347] In some embodiments, the composition or administration of the composition affects a measurement such as a liver fat percentage measurement, a liver fibrosis score measurement, a NAFLD activity score measurement, a blood alanine aminotransferase (ALT) measurement, a blood aspartate aminotransferase (AST) measurement, a blood alkaline phosphatase (ALP) measurement, a blood bilirubin measurement, a low-density lipoprotein (LDL) measurement, a total cholesterol measurement, a non-HDL cholesterol measurement, an apolipoprotein B (APOB), a GPAM protein measurement, or a GPAM mRNA measurement, relative to the baseline measurement.

[0348] Some embodiments of the methods described herein include obtaining the measurement from a subject. For example, the measurement may be obtained from the subject after treating the subject. In some embodiments, the measurement is obtained in a second sample (such as a fluid or tissue sample described herein) obtained from the subject after the composition is administered to the subject. In some embodiments, the measurement is an indication that the disorder has been treated.

[0349] In some embodiments, the measurement is obtained directly from the subject. In some embodiments, the measurement is obtained noninvasively using an imaging device. In some embodiments, the measurement is obtained in a second sample from the subject. In some embodiments, the measurement is obtained in one or more histological tissue sections. In some embodiments, the measurement is obtained by performing an assay on the second sample obtained from the subject. In some embodiments, the measurement is obtained by an assay, such as an assay described herein. In some embodiments, the assay is an immunoassay, a colorimetric assay, a fluorescence assay, a chromatography (e.g., HPLC) assay, or aPCR assay. In some embodiments, the measurement is obtained by an assay such as an immunoassay, a colorimetric assay, a fluorescence assay, or a chromatography (e.g., HPLC) assay. In some embodiments, the measurement is obtained by PCR. In some embodiments, the measurement is obtained by histology. In some embodiments, the measurement is obtained by observation. In some embodiments, additional measurements are made, such as in a 3rd sample, a 4th sample, or a fifth sample. [0350] In some embodiments, the measurement is obtained within 1 hour, within 2 hours, within 3 hours, within 4 hours, within 5 hours, within 6 hours, within 12 hours, within 18 hours, or within 24 hours after the administration of the composition. In some embodiments, the measurement is obtained within 1 day, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, or within 7 days after the administration of the composition. In some embodiments, the measurement is obtained within 1 week, within 2 weeks, within 3 weeks, within 1 month, within 2 months, within 3 months, within 6 months, within 1 year, within 2 years, within 3 years, within 4 years, or within 5 years after the administration of the composition. In some embodiments, the measurement is obtained after 1 hour, after 2 hours, after 3 hours, after 4 hours, after 5 hours, after 6 hours, after 12 hours, after 18 hours, or after 24 hours after the administration of the composition. In some embodiments, the measurement is obtained after 1 day, after 2 days, after 3 days, after 4 days, after 5 days, after 6 days, or after 7 days after the administration of the composition. In some embodiments, the measurement is obtained after 1 week, after 2 weeks, after 3 weeks, after 1 month, after 2 months, after 3 months, after 6 months, after 1 year, after 2 years, after 3 years, after 4 years, or after 5 years, following the administration of the composition.

[0351] In some embodiments, the composition improves the measurement relative to the baseline measurement. For example, an adverse phenotype of NAFLD, NASH, alcoholic liver disease, liver fibrosis, liver cirrhosis, hepatocellular carcinoma, hyperlipidemia, ischemic heart disease, or coronary heart disease may be improved upon administration of the composition. In some embodiments, the improvement is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the improvement is measured directly in the subject after administering the composition to the subject. In some embodiments, the measurement is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by about 10% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 10%, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline measurement. In some embodiments, the measurement is improved by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by arange defined by any of the two aforementioned percentages. [0352] In some embodiments, the composition improves the measurement relative to the baseline measurement. For example, a protective liver function phenotype, metabolic phenotype, or level of circulating ketone bodies may be improved upon administration of the composition. In some embodiments, the improvement is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the improvement is measured directly in the subject after administering the composition to the subject. In some embodiments, the measurement is improved by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by about 10% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by about 100% or more, improved by about 250% or more, improved by about 500% or more, improved by about 750% or more, or improved by about 1000% or more, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 2.5%, no more than about 5%, orno more than about 7.5%, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 10%, relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline measurement. In some embodiments, the measurement is improved by no more than about 100%, improved by no more than about 250%, improved by no more than about 500%, improved by no more than about 750%, or improved by no more than about 1000%, relative to the baseline measurement. In some embodiments, the measurement is improved by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by arange defined by any of the two aforementioned percentages.

[0353] In some embodiments, the measurement is a liver fat percentage (LFP) measurement. In some embodiments, the LFP measurement is a percentage as measured by MRI. In some embodiments, the LFP measurement is a percentage as measured by CT scan. In some embodiments, the LFP is measured in a liver biopsy.

[0354] In some embodiments, the composition reduces the LFP measurement relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by about 10% or more, relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by no more than about 10%, relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline LFP measurement. In some embodiments, the LFP measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0355] In some embodiments, the measurement is a liver fibrosis measurement. In some embodiments, the liver fibrosis measurement is a liver fibrosis score (LFS). In some embodiments, the LFS comprises a score of 0, 1, 2, 3, or 4, or a range of scores defined by any two of the aforementioned numbers. In some embodiments, the LFS comprises a score of 0-4. In some embodiments, the LFS is obtained using a scoring system exemplified in Table 2. In some embodiments, the LFS measurement is obtained noninvasively. In some embodiments, the LFS measurement is obtained by a medical imaging device such as a vibration-controlled transient elastography (VCTE) device, a shear wave elastography device, a medical resonance imaging (MRI) device, a magnetic resonance spectroscopy device, a computed tomography device, or an ultrasound device. In some embodiments, the LFS measurement is obtained in a liver sample. In some embodiments, the LFS is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. In some embodiments, the LFS is obtained using one or more indirect markers or measures of liver fibrosis such as an aspartate aminotransferase-to-platelet ratio index (APRI), a Fibrosis-4 (FIB-4) index, a FibroIndex, a Forns Index, a Hepascore, or a FibroTest. In some embodiments, the LFS is obtained using one or more indirect markers or measures of liver fibrosis such as a FIBROSpect test or a FIBROSpect II test. In some embodiments, the LFS is obtained by RT-qPCR or RNA sequencing of one or more fibrosis-related genes such as a collagen gene. In some embodiments, the LFS or the LFS is obtained using a scoring system upon a visual inspection of a sample such as a histological sample. In some embodiments, the LFS or the LFS is obtained using a stain with an affinity to collagen. In some embodiments, the LFS measurement is measured by FibroScan® in kilopascals (kPa). In some embodiments, the LFS is measured by histological analysis. In some embodiments, the LFS measurement is a LFS measurement assayed by imaging. In some embodiments, the imaging is by MRI or CT scan.

[0356] In some embodiments, the composition reduces the LFS measurement relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by about 10% or more, relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by no more than about 10%, relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline LFS measurement. In some embodiments, the LFS measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0357] In some embodiments, the measurement is a NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is measured by histological analysis. In some embodiments, the NAFLD activity score measurement includes a numerical score for steatosis (0-3), hepatocyte ballooning (1-2), and lobular inflammation (0-3). In some embodiments, the score thresholds of < 3 correlates with a diagnosis of not-NASH. In some embodiments, the score thresholds of > 5 correlates with a diagnosis of NASH. In some embodiments, a NAFLD activity score measurement is assayed by imaging.

[0358] In some embodiments, the composition reduces the NAFLD activity score measurement relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by about 10% or more, relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by no more than about 10%, relative to the baseline NAFLD activity score measurement. In some embodiments, the NAFLD activity score measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline NAFLD ACTIVITY SCORE measurement. In some embodiments, the NAFLD activity score measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages. [0359] In some embodiments, the measurement is an alanine aminotransferase (ALT) measurement. In some embodiments, the ALT measurement includes a blood ALT measurement. In some embodiments, the ALT measurement is measured by enzymatic activity. In some embodiments, the ALT measurement is measured by presence of an ALT epitope. In some embodiments, the ALT measurement is a circulating ALT measurement. In some embodiments, the ALT measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. In some embodiments, the ALT measurement is a blood ALT concentration (for example, units per liter (U/L)). In some embodiments, the ALT measurement is a circulating ALT measurement. In some embodiments, the ALT measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0360] In some embodiments, the composition reduces the ALT measurement relative to the baseline ALT measurement. In some embodiments, the composition reduces circulating ALT relative to the baseline ALT measurement. In some embodiments, the reduced ALT is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the ALT measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by about 10% or more, relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by no more than about 10%, relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline ALT measurement. In some embodiments, the ALT measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by arange defined by any of the two aforementioned percentages.

[0361] In some embodiments, the measurement is an aspartate aminotransferase (AST) measurement. In some embodiments, the AST measurement includes a blood AST measurement. In some embodiments, the AST measurement is measured by enzymatic activity. In some embodiments, the AST measurement is measured by presence of an AST epitope. In some embodiments, the AST measurement is a blood AST concentration (for example, units per liter (U/L)). In some embodiments, the AST measurement is a circulating AST measurement. In some embodiments, the AST measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay. [0362] In some embodiments, the composition reduces the AST measurement relative to the baseline AST measurement. In some embodiments, the composition reduces circulating AST relative to the baseline AST measurement. In some embodiments, the reduced AST is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the AST measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by about 10% or more, relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by no more than about 10%, relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline AST measurement. In some embodiments, the AST measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by arange defined by any of the two aforementioned percentages.

[0363] In some embodiments, the measurement is an alkaline phosphatase (ALP) measurement. In some embodiments, the ALP measurement includes a blood ALP measurement. In some embodiments, the ALP measurement is a blood ALP concentration (for example, units per liter (U/L)). In some embodiments, the ALP measurement is measured by enzymatic activity. In some embodiments, the ALP measurement is measured by presence of an ALP epitope. In some embodiments, the ALP measurement is a circulating ALP measurement. In some embodiments, the ALP measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0364] In some embodiments, the composition reduces the ALP measurement relative to the baseline ALP measurement. In some embodiments, the composition reduces circulating ALP relative to the baseline ALP measurement. In some embodiments, the reduced ALP is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the ALP measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by about 10% or more, relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by no more than about 10%, relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline ALP measurement. In some embodiments, the ALP measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by arange defined by any of the two aforementioned percentages.

[0365] In some embodiments, the measurement is a bilirubin measurement. In some embodiments, the bilirubin measurement includes a blood bilirubin measurement. In some embodiments, the bilirubin measurement is a blood bilirubin concentration (for example, units per liter (pmol/L)). In some embodiments, the bilirubin measurement is measured by presence of a bilirubin epitope. In some embodiments, the bilirubin measurement is a circulating bilirubin measurement. In some embodiments, the bilirubin measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0366] In some embodiments, the composition reduces the bilirubin measurement relative to the baseline bilirubin measurement. In some embodiments, the composition reduces circulating bilirubin relative to the baseline bilirubin measurement. In some embodiments, the reduced bilirubin is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the bilirubin measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by about 10% or more, relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by no more than about 10%, relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline bilirubin measurement. In some embodiments, the bilirubin measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages. [0367] In some embodiments, the measurement is a low-density lipoprotein (LDL) measurement. In some embodiments, the LDL measurement includes a blood LDL measurement. In some embodiments, the LDL measurement is a blood LDL concentration (for example, units per liter (mg/dL)). In some embodiments, the LDL measurement is measured by presence of an LDL epitope. In some embodiments, the LDL measurement is a circulating LDL measurement. In some embodiments, the LDL measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0368] In some embodiments, the composition reduces the LDL measurement relative to the baseline LDL measurement. In some embodiments, the composition reduces circulating LDLs relative to the baseline LDL measurement. In some embodiments, the reduced LDL is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the LDL measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by about 10% or more, relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by no more than about 10%, relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline LDL measurement. In some embodiments, the LDL measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by arange defined by any of the two aforementioned percentages.

[0369] In some embodiments, the measurement is an APOB measurement. In some embodiments, the APOB measurement is a blood APOB concentration (for example, units per liter (mg/dL)). In some embodiments, the APOB measurement is a circulating APOB measurement. In some embodiments, the APOB measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0370] In some embodiments, the composition reduces the APOB measurement relative to the baseline APOB measurement. In some embodiments, the composition reduces circulating APOBs relative to the baseline APOB measurement. In some embodiments, the reduced APOB is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the APOB measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by about 10% or more, relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by no more than about 10%, relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline APOB measurement. In some embodiments, the APOB measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0371] In some embodiments, the measurement is a total cholesterol measurement. In some embodiments, the total cholesterol measurement is a blood total cholesterol concentration (for example, units per liter (mg/dL)). In some embodiments, the total cholesterol measurement is a circulating total cholesterol measurement. In some embodiments, the total cholesterol measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0372] In some embodiments, the composition reduces the total cholesterol measurement relative to the baseline total cholesterol measurement. In some embodiments, the composition reduces circulating total cholesterol relative to the baseline total cholesterol measurement. In some embodiments, the reduced total cholesterol is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the total cholesterol measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by about 10% or more, relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by no more than about 10%, relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline total cholesterol measurement. In some embodiments, the total cholesterol measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0373] In some embodiments, the measurement is a non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is a blood non-HDL cholesterol concentration (for example, units per liter (mg/dL)). In some embodiments, the non-HDL cholesterol measurement is a circulating non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0374] In some embodiments, the composition reduces the non-HDL cholesterol measurement relative to the baseline non-HDL cholesterol measurement. In some embodiments, the composition reduces circulating non-HDL cholesterol relative to the baseline non-HDL cholesterol measurement. In some embodiments, the reduced non-HDL cholesterol is measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the non- HDL cholesterol measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non- HDL cholesterol measurement is decreased by about 10% or more, relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is decreased by no more than about 10%, relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or about 100% relative to the baseline non-HDL cholesterol measurement. In some embodiments, the non-HDL cholesterol measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0375] In some embodiments, the measurement is an GPAM protein measurement. In some embodiments, the GPAM protein measurement comprises an GPAM protein level. In some embodiments, the GPAM protein level is indicated as a mass or percentage of GPAM protein per sample weight. In some embodiments, the GPAM protein level is indicated as a mass or percentage of GPAM protein per sample volume. In some embodiments, the GPAM protein level is indicated as a mass or percentage of GPAM protein per total protein within the sample. In some embodiments, the GPAM protein measurement is a tissue GPAM protein measurement. In some embodiments, the GPAM protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.

[0376] In some embodiments, the composition reduces the GPAM protein measurement relative to the baseline GPAM protein measurement. In some embodiments, the composition reduces tissue GPAM protein levels relative to the baseline GPAM protein measurement. In some embodiments, the reduced GPAM protein levels are measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the GPAM protein measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by about 10% or more, relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by no more than about 10%, relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline GPAM protein measurement. In some embodiments, the GPAM protein measurement is decreased by 2.5%, 5%, 7.5%, 19%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.

[0377] In some embodiments, the measurement is an GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement comprises an GPAM mRNA level. In some embodiments, the GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per sample weight. In some embodiments, the GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per sample volume. In some embodiments, the GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per total mRNA within the sample. In some embodiments, the GPAM mRNA level is indicated as an amount or percentage of GPAM mRNA per total nucleic acids within the sample. In some embodiments, the GPAM mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the GPAM mRNA measurement is a tissue GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is obtained by an assay such as a PCR assay. In some embodiments, the PCR comprises qPCR. In some embodiments, the PCR comprises reverse transcription of the GPAM mRNA. [0378] In some embodiments, the composition reduces the GPAM mRNA measurement relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is obtained in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the composition reduces GPAM mRNA levels relative to the baseline GPAM mRNA levels. In some embodiments, the reduced GPAM mRNA levels are measured in a second sample obtained from the subject after administering the composition to the subject. In some embodiments, the second sample is a liver sample. In some embodiments, the second sample is an adipose sample. In some embodiments, the GPAM mRNA measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by about 10% or more, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by no more than about 10%, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, relative to the baseline GPAM mRNA measurement. In some embodiments, the GPAM mRNA measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% or by a range defined by any of the two aforementioned percentages.

III. DEFINITIONS

[0379] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0380] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0381] As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a sample” includes a plurality of samples, including mixtures thereof.

[0382] The terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement. The terms include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of’ can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.

[0383] The terms “subject,” and “patient” may be used interchangeably herein. A “subject” can be a biological entity containing expressed genetic materials. The biological entity can be a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The subject can be a mammal. The mammal can be a human. The subject may be diagnosed or suspected of being at high risk for a disease. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0384] As used herein, the term “about” a number refers to that number plus or minus 10% of that number. The term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.

[0385] As used herein, the terms “treatment” or “treating” are used in reference to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and/or a prophylactic benefit. A therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.

[0386] The term “C_x-y” or “C_x-C_y” when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain. For example, the term “C_1-6alkyl” refers to substituted or unsubstituted saturated hydrocarbon groups, including straight- chain alkyl and branched- chain alkyl groups that contain from 1 to 6 carbons. [0387] The terms “C_x-yalkenyl” and “C_x-yalkynyl” refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.

[0388] The term “carbocycle” as used herein refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. Carbocycle includes 3- to 10-membered monocyclic rings, 5- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. A bicyclic carbocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits. A bicyclic carbocycle further includes spiro bicyclic rings such as spiropentane. A bicyclic carbocycle includes any combination of ring sizes such as 3-3 spiro ring systems, 4-4 spiro ring systems, 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, naphthyl, and bicyclo[ 1.1. 1] pentanyl.

[0389] The term “aryl” refers to an aromatic monocyclic or aromatic multicyclic hydrocarbon ring system. The aromatic monocyclic or aromatic multicyclic hydrocarbon ring system contains only hydrogen and carbon and from five to eighteen carbon atoms, where at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) a -electron system in accordance with the Hückel theory. The ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene.

[0390] The term "cycloalkyl" refers to a saturated ring in which each atom of the ring is carbon. Cycloalkyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 5- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings. In certain embodiments, a cycloalkyl comprises three to ten carbon atoms. In other embodiments, a cycloalkyl comprises five to seven carbon atoms. The cycloalkyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, spiropentane, norbornyl (i.e., bicyclo[2.2. 1]heptanyl), decalinyl, 7,7-dimethyl bicyclo[2.2.1]heptanyl, bicyclo[1. 1.1]pentanyl, and the like.

[0391] The term "cycloalkenyl" refers to a saturated ring in which each atom of the ring is carbon and there is at least one double bond between two ring carbons. Cycloalkenyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 5- to 12-membered bridged rings. In other embodiments, a cycloalkenyl comprises five to seven carbon atoms. The cycloalkenyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0392] The term “halo” or, alternatively, “halogen” or “halide,” means fluoro, chloro, bromo or iodo. In some embodiments, halo is fluoro, chloro, or bromo.

[0393] The term “haloalkyl” refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1- chloromethyl-2-fluoroethyl, and the like. In some embodiments, the alkyl part of the haloalkyl radical is optionally further substituted as described herein.

[0394] The term “heterocycle” as used herein refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings. A bicyclic heterocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits. In an exemplary embodiment, an aromatic ring, e.g., pyridyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, morpholine, piperidine or cyclohexene. A bicyclic heterocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. A bicyclic heterocycle further includes spiro bicyclic rings, e.g., 5 to 12-membered spiro bicycles, such as 2-oxa-6-azaspiro[3.3]heptane.

[0395] The term "heteroaryl" refers to a radical derived from a 5 to 18 membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, the heteroaryl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) π-electron system in accordance with the Hückel theory. Heteroaryl includes fused or bridged ring systems. The heteroatom(s) in the heteroaryl radical is optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heteroaryl is attached to the rest of the molecule through any atom of the ring(s). Examples of heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3- benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo [b][ 1,4] oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl-(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H- cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6- dihy drobenzo| h | cinnolinyl, 6,7-dihydro-5H-benzo[6,7] cyclohepta[ 1 ,2-c] pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8- tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1 -phenyl- 1H- pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4- d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6, 7, 8, 9- tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2- d]pyrimidinyl, thieno[2,3-c]pyridinyl, and thiophenyl (i.e. thienyl).

[0396] The term "heterocycloalkyl" refers to a saturated ring with carbon atoms and at least one heteroatom. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycloalkyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12- membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings. The heteroatoms in the heterocycloalkyl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heterocycloalkyl is attached to the rest of the molecule through any atom of the heterocycloalkyl, valence permitting, such as any carbon or nitrogen atoms of the heterocycloalkyl. Examples of heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2- oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4- piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydro furyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1 -oxo-thiomorpholinyl, 2-oxa-6- azaspiro[3.3]heptane, and 1,1-dioxo-thiomorpholinyl.

[0397] The term "heterocycloalkenyl" refers to an unsaturated ring with carbon atoms and at least one heteroatom and there is at least one double bond between two ring carbons. Heterocycloalkenyl does not include heteroaryl rings. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycloalkenyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 5- to 12-membered bridged rings. In other embodiments, a heterocycloalkenyl comprises five to seven ring atoms. The heterocycloalkenyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, e.g., pyrroline (dihydropyrrole), pyrazoline (dihydropyrazole), imidazoline (dihydroimidazole), triazoline (dihydrotriazole), dihydrofuran, dihydrothiophene, oxazoline (dihydrooxazole), isoxazoline (dihydroisoxazole), thiazoline (dihydrothiazole), isothiazoline (dihydroisothiazole), oxadiazoline (dihydrooxadiazole), thiadiazoline (dihydrothiadiazole), dihydropyridine, tetrahydropyridine, dihydropyridazine, tetrahydropyridazine, dihydropyrimidine, tetrahydropyrimidine, dihydropyrazine, tetrahydropyrazine, pyran, dihydropyran, thiopyran, dihydrothiopyran, dioxine, dihydrodioxine, oxazine, dihydrooxazine, thiazine, and dihydrothiazine. [0398] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., anNH or NH₂ of a compound. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds.

[0399] In some embodiments, substituents may include any substituents described herein, for example: halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), oximo (=N-OH), hydrazino (=N-NH₂), -R^bOR^a, -R^bOC(O)R^a, -R^bOC(O)OR^a, -R^bOC(O)N(R^a)₂, -R^bN(R^a)₂, - R^bC(O)R^a, -R^bC(O)OR^a, -R^bC(O)N(R^a)₂, -R^bOR^cC(O)N(R^a)₂, -R^bN(R^a)C(O)OR^a, -R^bN(R^a)C(O)R^a, - R^bN(R^a)S(O)_tR^a (where t is 1 or 2), -R^bS(O)tR^a (where t is 1 or 2), -R^bS(O)_tOR^a (where t is 1 or 2), and -R^bS(O)tN(R^a)₂ (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, and heteroarylalkyl, any of which may be optionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), oximo (=N-OH), hydrazine (=N-NH₂), -R^bOR^a, -R^bOC(O)R^a, -R^bOC(O)OR^a, -R^bOC(O)N(R^a)₂, -R^bN(R^a)₂, -R^bC(O)R^a, - R^bC(O)OR^a, -R^bC(O)N(R^a)₂, -R^bOR^cC(O)N(R^a)₂, -R^bN(R^a)C(O)OR^a, -R^bN(R^a)C(O)R^a, - R^bN(R^a)S(O)_tR^a (where t is 1 or 2), -R^bS(O)tR^a (where t is 1 or 2), -R^bS(O)_tOR^a (where t is 1 or 2) and -R^bS(O)_tN(R^a)₂ (where t is 1 or 2); wherein each R^a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each R^a, valence permitting, may be optionally substituted with alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), oximo (=N-OH), hydrazine (=N-NH₂), -R^bOR^a, -R^bOC(O)R^a, - R^bOC(O)OR^a, -R^bOC(O)N(R^a)₂, -R^bN(R^a)₂, -R^bC(O)R^a, -R^bC(O)OR^a, -R^bC(O)N(R^a)₂, - R^bOR^cC(O)N(R^a)₂, -R^bN(R^a)C(O)OR^a, -R^bN(R^a)C(O)R^a, -R^bN(R^a)S(O)_tR^a (where t is 1 or 2), - R^bS(O)_tR^a (where t is 1 or 2), -R^bS(O)_tOR^a (where t is 1 or 2) and -R^bS(O)_tN(R^a)2 (where t is 1 or 2); and wherein each R^b is independently selected from a direct bond or a straight or branched alkylene, alkenylene, or alkynylene chain, and each R^c is a straight or branched alkylene, alkenylene or alkynylene chain.

[0400] Double bonds to oxygen atoms, such as oxo groups, are represented herein as both “=O” and “(O)” Double bonds to nitrogen atoms are represented as both “=NR” and “(NR)”. Double bonds to sulfur atoms are represented as both “=S” and “(S)”

[0401] In some embodiments, a "derivative" polypeptide or peptide is one that is modified, for example, by glycosylation, pegylation, phosphorylation, sulfation, reduction/alkylation, acylation, chemical coupling, or mild formalin treatment. A derivative may also be modified to contain a detectable label, either directly or indirectly, including, but not limited to, a radioisotope, fluorescent, and enzyme label.

[0402] Some embodiments refer to nucleic acid sequence information. It is contemplated that in some embodiments, thymine (T) may be interchanged with uracil (U), or vice versa. For example, some sequences in the sequence listing may recite Ts, but these may be replaced with Us in some embodiments. In some oligonucleotides with nucleic acid sequences that include uracil, the uracil ma^r be replaced with thymine. Similarly, in some oligonucleotides with nucleic acid sequences that include thymine, the thymine may be replaced with uracil. In some embodiments, an oligonucleotide such as an siRNA comprises or consists of RNA. In some embodiments, the oligonucleotide may include DNA. For example, the oligonucleotide may include 2’ deoxyribonucleotides. An ASO may comprise or consist of DNA. To any extent that the sequence listing contradicts the disclosure in the specification, the specification takes precedence.

[0403] Some aspects include sequences with nucleotide modifications or modified internucleoside linkages. Generally, and unless otherwise specified, Nf (e.g., Af, Cf, Gf, Tf, or Uf) refers to a 2’- fluoro-modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) refers to a 2’ deoxy nucleoside, n (e.g., a, c, g, t, or u) refers to a 2’-O-methyl modified nucleoside, and “s” refers to a phosphorothioate linkage.

[0404] A pyrimidine may include cytosine (C), thymine (T), or uracil (U). A pyrimidine may include C or U. A pyrimidine may include C or T. A reference to a pyrimidine may include a nucleoside or nucleotide comprising the pyrimidine. A purine may include guanine (G), inosine (I) or adenine (A). A reference to a purine may include a nucleoside or nucleotide comprising a purine.

[0405] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. IV. EXAMPLES

Example 1: Functional Variants in GPAM Demonstrate Protective Associations for Non- Alcoholic Fatty Liver Disease, Alcoholic Liver Disease, Decreased Liver Fat Percentage and Decreased Blood Lipids

[0406] Variants in GPAM were evaluated for associations with liver disease, liver fat percentage, liver function parameters, and blood lipid levels in approximately 452,000 individuals from the UK Biobank cohort. Variants evaluated included rs2792751, a common (AAF=0.73) missense variant (I43V); and rs57128949 a common (AAF=0. 18) intergenic variant that is a GPAM mRNA expression quantitative trait locus (eQTL) associated with decreased GPAM mRNA expression in multiple tissues, including liver. Stepwise conditional analyses in multiple traits, as well as direct evaluation of linkage disequilibrium, confirmed that rs2792751 and rs57128949 are independent variants. Some details of these variants are shown in Table 3.

Table 3. GPAM variants

[0407] The analyses resulted in identification of associations for the individual GPAM variants and phenotypes. Applicant hypothesizes that individually these variants result in a decrease in the abundance and/or activity of the GPAM gene product, and that it is this loss of function that leads to the observed genetic associations. This hypothesis is supported by associations between the GPAM variants and increased levels of blood ketone bodies as measured by nuclear magnetic resonance in the UK Biobank, including 3 -hydroxy butyrate, acetoacetate and acetone (Table 4). These results were consistent with observations in GPAM knockout mice, which demonstrate increased plasma ketone bodies, and with the known function of GPAM in triacylglycerol (TAG) metabolism.

Table 4. GPAM variant blood ketone associations

[0408] The analyses resulted in identification of associations for the GPAM variants. For example, there were protective associations with multiple liver disease-related traits. GPAM variants are individually associated with decreased risk of non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, liver fibrosis and cirrhosis, decreased risk of sequelae of chronic liver disease such as esophageal varices and portal hypertension, and with decreased MRI -derived liver fat percentage (Table 5) GPAM variants are also associated with decreased blood levels of liver enzymes, including alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), and with decreased blood bilirubin (Table 6).

Table 5. GPAM variant liver disease-related associations

Table 6. GPAM variant liver function associations

[0409] GPAM variants are also associated with decreased LDL-cholesterol, total cholesterol and APOB, and with decreased use of ‘statin’ (HMG CoA reductase inhibitor) medications (Table 7). These protective associations with putative loss of function variants in GPAM across several related and distinct diseases and traits suggest that inhibition of GPAM could be therapeutic in these and related diseases.

Table 7. GPAM variant blood lipid associations

Example 2: Bioinformatic selection of sequences in order to identify therapeutic siRNAs to downmodulate expression of the GPAM mRNA

[0410] Screening sets were defined based on bioinformatic analysis. Therapeutic siRNAs were designed to target human GPAM. Predicted specificity in human, rhesus monkey, cynomolgus monkey, mouse, rat, rabbit, and dog was determined for sense (S) and antisense (AS) strands. These were assigned a “specificity score” which considers the likelihood of unintended downregulation of any other transcript by full or partial complementarity of an siRNA strand (up to 2 mismatches within positions 2-18) as well as the number and positions of mismatches. Thus, off-target(s) transcripts for antisense and sense strands of each siRNA were identified. As identified, siRNAs with high specificity and a low number of predicted off-targets provided a benefit of increased targeting specificity.

[0411] In addition to selecting siRNA sequences with high sequence specificity to GPAM mRNA, siRNA sequences within the seed region were analyzed for similarity to seed regions of known miRNAs. siRNAs can function in a miRNA like manner via base-pairing with complementary sequences within the 3’-UTR of mRNA molecules. The complementarity typically encompasses the 5‘-bases at positions 2-7 of the miRNA (seed region). To circumvent siRNAs to act via functional miRNA binding sites, siRNA strands containing natural miRNA seed regions can be avoided. Seed regions identified in miRNAs from human, mouse, rat, rhesus monkey, dog, rabbit, and pig are referred to as “conserved”. Combining the “specificity score” with miRNA seed analysis yielded a “specificity category”. This is divided into categories 1-4, with 1 having the highest specificity and 4 having the lowest specificity. Each strand of the siRNA is assigned to a specificity category.

[0412] Analysis of the Genome Aggregation Database (gnomAD) to identify siRNAs targeting regions with known SNPs was also carried out to identify siRNAs that may be non -functional in individuals containing the SNP. Information regarding the positions of SNPs within the target sequence as well as minor allele frequency (MAF) in case data was obtained in this analysis.

[0413] Initial analysis of the relevant GPAM mRNA sequence revealed few sequences that fulfil the specificity parameters and at the same time target GPAM mRNA in all the analyzed relevant species. Therefore, independent screening subsets were designed for the therapeutic siRNAs.

[0414] The siRNAs in these subsets recognized at least the human GPAM sequences. Therefore, the siRNAs in these subsets can be used to target human GPAM in a therapeutic setting.

[0415] The number of siRNA sequences derived from human GPAM mRNA (ENST00000348367.9, SEQ ID NO: 12867) without consideration of specificity or species cross-reactivity was 6354 (sense and antisense strand sequences included in SEQ ID NOS: 1 -6354 and 6355-12708, respectively) as shown in Table 8.

[0416] Prioritizing sequences for target specificity, miRNA seed region sequences and SNPs as described above yields subset A. Subset A contains 1348 siRNAs whose base sequences are shown in Table 8.

[0417] The above methods can be used to identify therapeutic siRNAs to downmodulate expression of the GPAM mRNA.

Table 8. Sequences in siRNA subset A- J

[0418] The siRNAs in subset A had the following characteristics: Cross -reactivity: With 19mer in human GPAM mRNA; Specificity category: For human: AS2 or better, SS3 or better; and miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species; Off-target frequency: ≤30 human off-targets matched with 2 mismatches in antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18.)

[0419] The siRNA sequences in subset A were selected for more stringent specificity to yield subset B. Subset B included the 1344 siRNAs as shown in Table 8.

[0420] The siRNAs in subset B had the following characteristics: Cross-reactivity: With 19mer in human GPAM mRNA; Specificity category: For human: AS2 or better, SS3 or better; miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species; Off-target frequency: ≤20 human off-targets matched with 2 mismatches in antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18).

[0421] The siRNA sequences in subset B were further selected for absence of seed regions in the AS strand that are identical to a seed region of known human miRNA to yield subset C. Subset C included the 931 siRNAs as shown in Table 8.

[0422] The siRNAs in subset C had the following characteristics: Cross-reactivity: With 19mer in human GPAM mRNA; Specificity category: For human: AS2 or better, SS3 or better; miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species. AS strand: seed region not identical to seed region of known human miRNA; Off-target frequency: ≤30 human off-targets matched with 2 mismatches by antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18).

[0423] The siRNA sequences in subset C were also selected for absence of seed regions in the AS or S strands that are identical to a seed region of known human miRNA in addition to having an off- target frequency of ≤30 human off-targets matched with 2 mismatches by antisense strand to yield subset D. Subset D included 595 siRNAs as shown in Table 8.

[0424] Any siRNA among any of subsets A-E may comprise any modification pattern described herein. If a sequence has a different number of nucleotides in length than a modification pattern, the modification pattern may still be used with the appropriate number of additional nucleotides added 5’ or 3’ to match the number of nucleotides in the modification pattern. For example, if a sense or antisense strand of the siRNA among any of subsets A-E comprises 19 nucleotides, and a modification pattern comprises 21 nucleotides, UU may be added onto the 5’ end of the sense or antisense strand.

[0425] Therapeutic siRNAs were designed to target human GPAM as described above and, in some cases, the GPAM sequence of at least one toxicology-relevant species, in this case, the non-human primate (NHP) cynomolgus monkey. The siRNAs included in subset F had the following characteristics: Cross-reactivity: With 19mer in human GPAM mRNA, with 17mer/19mer in NHP GPAM; Specificity category: For human and NHP: AS2 or better, SS3 or better. Subset F included 25 siRNAs whose base sequences are shown in Table 8.

[0426] In some cases, the sense strand of any of the siRNAs of subset F comprises siRNA with a particular modification pattern. In this modification pattern, position 9 counting from the 5’ end of the of the sense strand is has the 2’F modification. If position 9 of the sense strand is a pyrimidine, then all purines in the sense strand have the 2’OMe modification. If position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with the 2’F modification in the sense strand. If position 9 and only one other base between positions 5 and 11 of the sense strand are pyrimidines, then both of these pyrimidines are the only two positions with the 2’F modification in the sense strand. If position 9 and only two other bases between positions 5 and 11 of the sense strand are pyrimidines, and those two other pyrimidines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. If there are >2 pyrimidines between positions 5 and 11 of the sense strand, then all combinations of pyrimidines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that the sense strand does not have three 2’F modifications in a row.

[0427] If position 9 of the sense strand is a purine, then all purines in the sense strand have the 2’OMe modification. If position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with the 2’F modification in the sense strand. If position 9 and only one other base between positions 5 and 11 of the sense strand are purines, then both of these purines are the only two positions with the 2’F modification in the sense strand. If position 9 and only two other bases between positions 5 and 11 of the sense strand are purines, and those two other purines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total. If there are >2 purines between positions 5 and 11 of the sense strand, then all combinations of purines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that the sense strand does not have three 2’F modifications in a row.

[0428] In some cases, position 9 of the sense strand can be a 2’deoxy. In these cases, 2’F and 2’OMe modifications may occur at the other positions of the sense strand.

[0429] In some cases, the sense strand of any of the siRNAs of subset F comprises a modification pattern which conforms to these sense strand rules (Table 9).

[0430] In some cases, the antisense strand of any of the siRNAs of subset F comprise a modification or modification pattern. Some such examples are included in Table 9. The siRNAs in Table 9 include a GalNAc moiety. Table 10 includes some additional sense strand modifications of the siRNAs in subset F. The siRNAs in subset F may comprise any other modification pattem(s). In the tables, Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxy nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate linkage.

Table 9: Modified Screening Set (Subset G)

Table 10: Additional Examples of Modified siRNAs (Subset H)

[0431] The siRNAs targeting human GPAM can also be selected using other criteria including crossreactivity with the NHP cynomolgus GPAM, and in some cases mouse GPAM. Selection of these siRNAs yields Subset I. Subset I includes 25 siRNAs whose base sequences are shown in Table 8.

[0432] The siRNAs in subset I have the following characteristics:

• Cross-reactivity: With 19mer in human GPAM mRNA, with 17mer or 19mer in NHP GPAM mRNA, and in some cases 17mer or 19mer in mouse GPAM.

• Specificity category: For human and NHP only cross-reactive siRNAs: human AS1 or better, human SS3 or better. For human, NHP and mouse cross-reactive siRNAs, human AS2 or better, human AS3 or better.

• miRNA seeds: For human and NHP only cross-reactive siRNAs: AS and SS: seed region not in any human siRNA, seed region not conserved in human, mouse, and rat and not present in >3 species. For human, NHP and mouse cross-reactive siRNAs: seed region not in any human siRNA, seed region not conserved in human, mouse, and rat and not present in >1 species.

• SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18)

[0433] The siRNAs targeting human GPAM can also be selected using at least Selection Set C criteria, and including cross-reactivity with the NHP cynomolgus GPAM. Selection of these siRNAs yields Subset J. Subset J includes 24 siRNAs whose base sequences are shown in Table 8.

[0434] The siRNAs in subset J have the following characteristics:

• Cross-reactivity: With 19mer in human GPAM mRNA, with 17mer or 19mer in NHP GPAM mRNA.

• Specificity category: human AS2 or better, human SS3 or better. NHP AS2 or better, NHP SS3 or better

• miRNA seeds: For human and NHP only cross-reactive siRNAs: AS strand: seed region not in any human siRNA, seed region not conserved in human, mouse, and rat and not present in >4 species. SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species.

• SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18)

Example 3: Chemically modified GPAM siRNAs

[0435] Any set of modifications, or any modification pattern may be used. For example, the siRNAs targeting GPAM can be synthesized with chemical modifications with the sense strand having modification pattern IS and antisense strand having modification pattern 1AS. In some embodiments, the siRNAs targeting GPAM can also be synthesized with chemical modifications with the sense strand having modification pattern 2S and antisense strand having modification pattern 3 AS. In some embodiments, the modifications included in Table 10 or Table 11 are used. In addition, adenosine can be placed at position 19 in the sense strand and uridine at position 1 in the antisense strand. Example 4: siRNA-mediated knockdown of GPAM in human hepatocyte cells

[0436] siRNAs targeted to the GPAM mRNA that downregulate levels of GPAM mRNA may lead to subsequent decrease of lysophosphatidic acid (LPA), a metabolite produced by the GPAM gene product, when administered to cultured hepatocyte cells.

[0437] On Day 0, the hepatocyte cells are to be seeded at 150,000 cells/mL into a Falcon 24-well tissue culture plate (ThermoFisher Cat. No. 353047) at 0.5 mL per well.

[0438] On Day 1, the GPAM siRNA and negative control siRNA master mixes are prepared. The GPAM siRNA master mix contains 350 μL of Opti-MEM (ThermoFisher Cat. No. 4427037 - sl288 Lot No. AS02B02D) and 3.5 μL of a mixture of the two GPAM siRNAs (10 pM stock). The negative control siRNA master mix contains 350 μL of Opti-MEM and 3.5 μL of negative control siRNA (ThermoFisher Cat. No. 4390843, 10 pM stock). Next, 3 μL of TransIT-X2 (Minis Cat. No. MIR6000) is added to each master mix. The mixes are incubated for 15 minutes to allow transfection complexes to form, then 51 μL of the appropriate master mix + TransIT-X2 is added to duplicate wells of hepatocyte cells with a final siRNA concentration of 10 nM.

[0439] On Day 3, 48 hours post transfection, cells are lysed using the Cells-to-Ct kit according to the manufacturer’s protocol (ThermoFisher Cat. No. 4399002). In brief, cells are washed with 50 μL using cold IX PBS and lysed by adding 49.5 μL of Lysis Solution and 0.5 μL DNase I per well and pipetting up and down 5 times and incubating for 5 minutes at room temperature. The Stop Solution (5 μL/well) is added to each well and mixed by pipetting up and down five times and incubating at room temperature for 2 minutes. The reverse transcriptase reaction is performed using 22.5 μL of the lysate according to the manufacturer’s protocol. Samples are stored at -80 °C until real-time qPCR was performed in triplicate using TaqMan Gene Expression Assays (Applied Biosystems FAM/GPAM using a BioRad iCycler).

[0440] A decrease in GPAM mRNA expression in the hepatocyte cells is expected after transfection with the GPAM siRNAs compared to GPAM mRNA levels in hepatocyte cells transfected with the non-specific control siRNA 48 hours after transfection. There is an expected decrease in the amount of LPA in wells containing hepatocytes cells transfected with the GPAM siRNAs relative to the amount of LPA in wells containing hepatocyte cells transfected with a non-specific control siRNA 48 hours after transfection. These results show that the GPAM siRNAs elicit knockdown of GPAM mRNA in hepatocyte cells and that the decrease in GPAM expression is correlated with a decrease in LPA production.

[0441] The GPAM siRNAs showing the greatest degree of knockdown of GPAM mRNA at 10 nM will be tested in a second screen for activity at 1 nM concentration using the transfection procedures as described above. Example 5: siRNA-mediated knockdown of GPAM in human adipocyte cells

[0442] Adipose tissue is a major site of endogenous triglyceride synthesis. siRNAs targeted to the GPAM mRNA that downregulate levels of GPAM mRNA may lead to subsequent decrease of lysophosphatidic acid (LPA), a metabolite produced by the GPAM gene product, when administered to cultured adipocyte cells. A decrease in LPA production may lead to a subsequent decrease in the levels of other components of the glycerol phosphate pathway for de novo triacylglycerol synthesis. Namely, phosphatidic acid (PA), phosphatidylserine (DAG), and triacylglycerol (TAG) may be decreased after siRNA-mediated knockdown of GPAM in human adipocyte cells.

[0443] On Day 0, the differentiated adipocyte cells (Cell Applications, Inc.) are to be seeded at 150,000 cells/mL into a Falcon 24-well tissue culture plate (ThermoFisher Cat. No. 353047) at 0.5 mL per well and cultured in adipocyte maintenance medium according to manufacturer’s instructions. [0444] On Day 1, the GPAM siRNA and negative control siRNA master mixes are prepared. The GPAM siRNA master mix contains 350 μL of Opti-MEM (ThermoFisher Cat. No. 4427037 - sl288 Lot No. AS02B02D) and 3.5 μL of a mixture of the two GPAM siRNAs (10 pM stock). The negative control siRNA master mix contains 350 μL of Opti-MEM and 3.5 μL of negative control siRNA (ThermoFisher Cat. No. 4390843, 10 pM stock). Next, 3 μL of TransIT-X2 (Minis Cat. No.

MIR6000) is added to each master mix. The mixes are incubated for 15 minutes to allow transfection complexes to form, then 51 μL of the appropriate master mix + TransIT-X2 is added to duplicate wells of differentiated adipocyte cells with a final siRNA concentration of 10 nM.

[0445] On Day 3, 48 hours post transfection, cells are lysed using the Cells-to-Ct kit according to the manufacturer’s protocol (ThermoFisher Cat. No. 4399002). In brief, cells are washed with 50 μL using cold IX PBS and lysed by adding 49.5 μL of Lysis Solution and 0.5 μL DNase I per well and pipetting up and down 5 times and incubating for 5 minutes at room temperature. The Stop Solution (5 μL/well) is added to each well and mixed by pipetting up and down five times and incubating at room temperature for 2 minutes. The reverse transcriptase reaction is performed using 22.5 μL of the lysate according to the manufacturer’s protocol. Samples are stored at -80 °C until real-time qPCR was performed in triplicate using TaqMan Gene Expression Assays (Applied Biosystems FAM/GPAM using a BioRad iCycler).

[0446] A decrease in GPAM mRNA expression in the adipocyte cells is expected after transfection with the GPAM siRNAs compared to GPAM mRNA levels in adipocyte cells transfected with the non-specific control siRNA 48 hours after transfection. There is an expected decrease in the amount of LPA in wells containing adipocyte cells transfected with the GPAM siRNAs relative to the amount of LPA in wells containing adipocyte cells transfected with a non-specific control siRNA 48 hours after transfection. These results show that the GPAM siRNAs elicit knockdown of GPAM mRNA in adipocyte cells and that the decrease in GPAM expression is correlated with a decrease in LPA production. PA, DAG, and TAG levels are measured in wells containing adipocyte cells transfected with a GPAM siRNA and with a non-specific control siRNA 48 hours after transfection if there was compensation for reduced GPAM activity at various steps of the tri acylglycerol synthesis pathway. These results show that the GPAM siRNAs elicit knockdown of GPAM mRNA in adipocyte cells and that the decrease in GPAM expression is correlated with a decrease in LPA production, PA production, DAG production, and TAG production.

[0447] The GPAM siRNAs showing the greatest degree of knockdown of GPAM mRNA at 10 nM will be tested in a second screen for activity at 1 nM concentration using the transfection procedures as described above.

Example 6: ASO-mediated knockdown of GPAM in human hepatocyte cells

[0448] ASOs targeted to the GPAM mRNA that downregulate levels of GPAM mRNA leading to subsequent decrease of LPA, a metabolite produced by the GPAM gene product, when administered to cultured hepatocyte cells.

[0449] On Day 0, the hepatocyte cells are seeded at 150,000 cells/mL into a Falcon 24-well tissue culture plate (Cat. No. 353047) at 0.5 mL per well.

[0450] On Day 1, the GPAM ASO and negative control ASO master mixes are prepared. The GPAM ASO master mix contains 350 μL of Opti-MEM (ThermoFisher Cat. No. 4427037 - sl288 Lot No. AS02B02D) and 3.5 μL of a mixture of the two GPAM ASOs (10 pM stock). The negative control ASO master mix contains 350 μL of Opti-MEM and 3.5 μL of negative control ASO (ThermoFisher Cat. No. 4390843, 10 pM stock). Next, 3 μL of TransIT-X2 (Mirus Cat. No. MIR6000) is added to each master mix. The mixes are incubated for 15 minutes to allow transfection complexes to form, then 51 μL of the appropriate master mix + TransIT-X2 is added to duplicate wells of hepatocyte cells with a final ASO concentration of 10 nM.

[0451] On Day 3, 48 hours post transfection, cells are lysed using the Cells-to-Ct kit according to the manufacturer’s protocol (ThermoFisher Cat. No. 4399002). In brief, cells are washed with 50 μL using cold 1X PBS and lysed by adding 49.5 μL of Lysis Solution and 0.5 μL DNase I per well and pipetting up and down 5 times and incubating for 5 minutes at room temperature. The Stop Solution (5 μL/well) is added to each well and mixed by pipetting up and down five times and incubating at room temperature for 2 minutes. The reverse transcriptase reaction is performed using 22.5 μL of the lysate according to the manufacturer’s protocol. Samples are stored at -80 °C until real-time qPCR was performed in triplicate using TaqMan Gene Expression Assays (Applied Biosystems FAM/GPAM using a BioRad iCycler).

[0452] A decrease in GPAM mRNA expression in the hepatocyte cells is expected after transfection with the GPAM ASOs compared to GPAM mRNA levels in hepatocyte cells transfected with the nonspecific control ASO 48 hours after transfection. There is an expected decrease in the amount of LPA in wells containing hepatocytes cells transfected with the GPAM ASOs relative to the amount of LPA in wells containing hepatocyte cells transfected with a non-specific control ASO 48 hours after transfection. These results show that the GPAM ASOs elicit knockdown of GPAM mRNA in hepatocyte cells and that the decrease in GPAM expression is correlated with a decrease in LPA production.

Example 7: Inhibition of GPAM in a Mouse Model for Fatty Liver Disease Using Modified GPAM siRNAs and ASOs

[0453] A murine model of fatty liver disease is used to evaluate the effect of siRNA or ASO inhibition of GPAM. In the murine model, fatty liver disease is induced by feeding mice a Western Diet (WD) containing 21. 1% fat, 41% Sucrose, and 1.25% Cholesterol by weight (Teklad diets, TD. 120528) and a high sugar solution (23.1g/L d-fructose (Sigma- Aldrich, G8270) and 18.9 g/L d- glucose (Sigma-Aldrich, F0127)) for 12 weeks. At 4-week-old C57BL/6J mice are fed a Western Diet instead of regular chow for 12 weeks.

[0454] Briefly, mice are divided into five groups: Group 1 - a fatty liver disease group treated with non-targeting control siRNA, Group 2 - a fatty liver disease group treated with non -targeting control ASO, Group 3 - a fatty liver disease group treated with GPAM siRNAl, Group 4 - a fatty liver disease group treated with GPAM ASO1, Group 5 - control mice on a normal chow diet. Each group contains eight mice (4 males, 4 females).

[0455] At weeks 12 weeks of Western Diet, blood samples are collected from each group prior to the first treatment.

[0456] Administration of siRNA or ASO is achieved with a 200ul subcutaneous injection of siRNA or ASO resuspended in PBS at concentration of lOuM. On Study Day 0, Group 1 mice are injected subcutaneously with non-targeting control siRNA, Group 2 mice are injected subcutaneously with non-targeting control ASO, Group 3 mice are injected subcutaneously with siRNAl targeting mouse GPAM, Group 4 mice are injected subcutaneously with AS 01 targeting mouse GPAM, and Group 5 mice are injected subcutaneously with vehicle. Every other week thereafter starting on Day 14 the animals from each group are dosed as on Day 0 for a total of 5 injections.

[0457] Weekly blood draws are taken and serum and plasma isolated. ALT, AST, ALP bilirubin, total cholesterol, HDL cholesterol and triglyceride levels are measured using VITROS 5,1 FS (Ortho Clinical Diagnostics). Non-fasting plasma insulin is measured with the Ultrasensitive Mouse Insulin ELISA kit (Crystal Chem, 90080) according to the manufacturer’s instructions. Non-fasting blood glucose is assayed with the One Touch Ultra (Life Scan). HOMA IR and QUICKI are calculated. Blood ketones, including 3 -hydroxy butyrate, acetoacetate and acetone are measured using EnzyChrom Ketone Body Assay Kit (BioAssay Systems, EKBD-100).

[0458] At the end of 12 weeks of Western Diet and siRNA/ASO treatment, mice are sacrificed by cervical dislocation following an intraperitoneal injection of 0.3 ml Nembutal (5 mg/ml). Terminal blood draw is collected via cardiac puncture and final ALT, AST, ALP bilirubin, total cholesterol, HDL cholesterol and triglyceride levels are measured along with non-fasting plasma insulin and glucose and blood ketones. Livers and adipose tissue are removed and divided into three sections each; one section placed in RNAlater for mRNA isolation, one section flash-frozen for protein isolation, one section fixed in formalin and then paraffin-embedded.

[0459] mRNA is isolated from tissue placed in RNAlater solution using the PureLink kit according to the manufacturer’s protocol (ThermoFisher Cat. No. 12183020). The reverse transcriptase reaction is performed according to the manufacturer’s protocol. Samples are stored at -80 °C until real-time qPCR was performed in triplicate using TaqMan Gene Expression Assays (Applied Biosystems FAM/GPAM using a BioRad iCycler). A decrease in GPAM mRNA expression in the liver and adipose tissue from mice is dosed with the GPAM siRNAs and ASOs compared to GPAM mRNA levels in the liver and adipose tissue from mice is dosed with the non-specific control siRNA and ASO. There is an expected decrease in the amount of SDF-1 in the liver and adipose tissue from mice that receive the GPAM siRNAs and ASOs compared to the amount of SDF - 1 in the liver and adipose tissue from mice that receive the non-specific control siRNA or ASO. These results show that the GPAM siRNAs and ASOs elicit knockdown of GPAM mRNA in liver and adipose tissue and that the decrease in GPAM expression is correlated with a decrease in SDF - 1 production. There is an expected decrease in blood ALT, AST, ALP, bilirubin and total cholesterol, and an expected increase in blood ketones in mice that receive the GPAM siRNA or ASO compared to the blood ALT, AST, ALP, bilirubin, total cholesterol and ketones in mice that receive the non-specific control. These results show that the GPAM siRNA or ASO elicit knockdown of GPAM mRNA in liver and adipose tissue and that the decrease in GPAM expression is correlated with a decrease in blood ALT, AST, ALP, bilirubin and total cholesterol, and an increase in blood ketones.

[0460] Formalin- fixed, paraffin-embedded liver sections are stained with hematoxylin and eosin (H&E) for assessment of liver histology, with Sirius Red (Sigma, 365548 -5G)/Fast Green (Sigma, F258) for assessment of fibrosis, and with periodic acid-Schiff (PAS) for assessment of glycogen accumulation. NAFLD Activity Score (NAS) and fibrosis stage are evaluated by an expert pathologist according to the NASH CRN scoring system. The histological scoring is performed blinded, with no knowledge by the pathologist of the treatment(s) received. These results show that the GPAM siRNAs and ASOs elicit knockdown of GPAM mRNA in liver tissue and that the decrease in GPAM expression is correlated with a decrease in NAS and NASH CRN.

Example 8: Oligonucleotide Synthesis

[0461] Oligonucleotides such as siRNAs may be synthesized according to phosphoramidite technology on a solid phase. For example, a K&A oligonucleotide synthesizer may be used. Syntheses may be performed on a solid support made of controlled pore glass (CPG, 500 A or 600 A, obtained from AM Chemicals, Oceanside, CA, USA). All 2'-OMe and 2’-F phosphoramidites may be purchased fromHongene Biotech (Union City, CA, USA). All phosphoramidites may be dissolved in anhydrous acetonitrile (100 mM) and molecular sieves (3 A) may be added. 5 -Benzylthio- 1H- tetrazole (BTT, 250 mM in acetonitrile) or 5 -Ethylthio- IH-tetrazole (ETT, 250 mM in acetonitrile) may be used as activator solution. Coupling times may be 9-18 min (e.g., with a GalNAc such as ETL17), 6 min (e.g., with 2'0Me and 2'F). In order to introduce phosphorothioate linkages, a 100 mM solution of 3-phenyl l,2,4-dithiazoline-5-one (POS, obtained from PolyOrg, Inc., Leominster, Mass., USA) in anhydrous acetonitrile may be employed.

[0462] After solid phase synthesis, the dried solid support may be treated with a 1 : 1 volume solution of 40 wt. % methylamine in water and 28% ammonium hydroxide solution (Aldrich) for two hours at 30° C. The solution may be evaporated and the solid residue may be reconstituted in water and purified by anionic exchange HPLC using a IKSgel SuperQ-5PW 13u column. Buffer A may be 20 mM Tris, 5 mM EDTA, pH 9.0 and contained 20% Acetonitrile and buffer B may be the same as buffer A with the addition of 1 M sodium chloride. UV traces at 260 nm may be recorded.

Appropriate fractions may be pooled then desalted using Sephadex G-25 medium.

[0463] Equimolar amounts of sense and antisense strand may be combined to prepare a duplex. The duplex solution may be prepared in 0.1 xPBS (Phosphate-Buffered Saline, 1 x, Gibco). The duplex solution may be annealed at 95° C. for 5 min, and cooled to room temperature slowly. Duplex concentration may be determined by measuring the solution absorbance on a UV-Vis spectrometer at 260 nm in 0.1 xPBS. For some experiments, a conversion factor may be calculated from an experimentally determined extinction coefficient.

Example 9: GalNAc ligand for hepatocyte targeting of oligonucleotides

[0464] Without limiting the disclosure to these individual methods, there are at least two general methods for attachment of multivalent N-acetylgalactosamine (GalNAc) ligands to oligonucleotides: solid or solution-phase conjugations. GalNAc ligands may be attached to solid phase resin for 3’ conjugation or at the 5’ terminus using GalNAc phosphorami dite reagents. GalNAc phosphorami dites may be coupled on solid phase as for other nucleosides in the oligonucleotide sequence at any position in the sequence. Reagents for GalNAc conjugation to oligonucleotides are shown in

Table 11

Table 11. GalNAc Conjugation Reagents

[0465] In solution phase conjugation, the oligonucleotide sequence — including a reactive conjugation site — is formed on the resin. The oligonucleotide is then removed from the resin and GalNAc is conjugated to the reactive site.

[0466] The carboxy GalNAc derivatives may be coupled to amino-modified oligonucleotides. The peptide coupling conditions are known to the skilled in the art using a carbodiimide coupling agent like DCC (N,N'-Dicyclohexylcarbodiimide), EDC (N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide) or EDC.HCl (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and an additive like HOBt (1-hydroxybenztriazole), HOSu (N-hydroxy succinimide), TBTU (N,N,N',N'-Tetramethyl-O-(benzotriazol-l-yl)uronium tetrafluoroborate, HBTU (2-(1H-benzotriazol- l-yl)-l, 1, 3, 3-tetramethyluronium hexafluorophosphate) or HOAt (l-Hydroxy-7-azabenzotriazole and common combinations thereof such as TBTU/HOBt or HBTU/HOAt to form activated amine-reactive esters.

[0467] amine groups may be incorporated into oligonucleotides using a number of known, commercially available reagents at the 5’ terminus, 3’ terminus or anywhere in between.

[0468] Non-limiting examples of reagents for oligonucleotide synthesis to incorporate an amino group include:

• 5’ attachment: • 6-(4-Monomethoxytritylamino)hexyl-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite CAS Number: 114616-27-2

• 5'-amino-Modifier TEG CE-Phosphoramidite

• 10-(O-trifluoroacetamido-N-ethyl)-triethyleneglycol-l-[(2-cyanoethyl)-(N,N-diisopropyl)]- phosphoramidite

• 3’ attachment:

• 3'-amino-Modifier Serinol CPG

• 3-Dimethoxytrityloxy-2-(3-(fluorenylmethoxycarbonylamino)propanamido)propyl-l-O- succinyl-long chain alkylamino-CPG (where CPG stands for controlled-pore glass and is the solid support)

• amino-Modifier Serinol Phosphorami dite

• 3-Dimethoxytrityloxy-2-(3-(fluorenylmethoxycarbonylamino)propanamido)propyl-l-O-(2- cyanoethyl)-(N,N-diisopropyl)-phosphoramidite

[0469] Internal (base modified):

• amino-Modifier C6 dT

• 5'-Dimethoxytrityl-5-[N-(trifluoroacetylaminohexyl)-3-acrylimido]-2'-deoxyUridine,3'-[(2- cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite. CAS Number: 178925-21-8

[0470] Solution phase conjugations may occur after oligonucleotide synthesis via reactions between non-nucleosidic nucleophilic functional groups that are attached to the oligonucleotide and electrophilic GalNAc reagents. Examples of nucleophilic groups include amines and thiols, and examples of electrophilic reagents include activated esters (e.g., N-hydroxysuccinimide, pentafluorophenyl) and mal eimides.

Example 10: GalNAc ligands for hepatocyte targeting of oligonucleotides

[0471] Without limiting the disclosure to these individual methods, there are at least two general methods for attachment of multivalent N-acetylgalactosamine (GalNAc) ligands to oligonucleotides: solid or solution-phase conjugations. GalNAc ligands may be attached to solid phase resin for 3’ conjugation or at the 5’ terminus using GalNAc phosphorami dite reagents. GalNAc phosphorami dites may be coupled on solid phase as for other nucleosides in the oligonucleotide sequence at any position in the sequence. A non-limiting example of a phosphoramidite reagent for GalNAc conjugation to a 5’ end oligonucleotide is shown in Table 12. Table 12. GalNAc Conjugation Reagent

[0472] The following includes examples of synthesis reactions used to create a GalNAc moiety:

Scheme for the preparation of NAcegal-Linker-TMSOTf

General procedure for preparation of Compound 2A

[0473] To a solution of Compound 1 A (500 g, 4.76 mol, 476 mL) in 2-Methly-THF (2.00 L) is added CbzCI (406 g, 2.38 mol, 338 mL) in 2-Methyl-THF (750mL) dropwise at 0 °C. The mixture is stirred at 25 °C for 2 hrs under N₂ atmosphere. TLC (DCM: MeOH = 20: 1, PMA) may indicate CbzCI is consumed completely and one new spot (Rr = 0.43) formed. The reaction mixture is added HCl/EtOAc (I N, 180 mL) and stirred for 30 mins, white solid is removed by filtration through celite, the filtrate is concentrated under vacuum to give Compound 2A (540 g, 2.26 mol, 47.5% yield) as a pale yellow oil and used into the next step without further purification. ¹ H NMR: δ 7.28 - 7.41 (m, 5 H), 5.55 (br s, 1 H), 5.01 - 5.22 (m, 2 H), 3.63 - 3.80 (m, 2 H), 3.46 - 3.59 (m, 4 H), 3.29 - 3.44 (m, 2 H), 2.83 - 3.02 (m, 1 H).

General procedure for preparation of Compound 4A

[0474] To a solution of Compound 3 A (1.00 kg, 4.64 mol, HCl) in pyridine (5.00 L) is added acetyl acetate (4.73 kg, 46.4 mol, 4.34 L) dropwise at 0 °C under N₂ atmosphere. The mixture is stirred at 25 °C for 16 hrs under N₂ atmosphere. TLC (DCM: MeOH = 20: 1, PMA) indicated Compound 3A is consumed completely and two new spots (Rr = 0.35) formed. The reaction mixture is added to cold water (30.0 L) and stirred at 0 °C for 0.5 hr, white solid formed, filtered and dried to give Compound 4A (1.55 kg, 3.98 mol, 85.8% yield) as a white solid and used in the next step without further purification. ¹ H NMR: δ 7.90 (d, J = 9.29 Hz, 1 H), 5.64 (d, J = 8.78 Hz, 1 H), 5.26 (d, J = 3.01 Hz, 1 H), 5.06 (dd, J = 11.29, 3.26 Hz, 1 H), 4.22 (t, J = 6. 15 Hz, 1 H), 3.95 - 4. 16 (m, 3 H), 2.12 (s, 3 H), 2.03 (s, 3 H), 1.99 (s, 3 H), 1.90 (s, 3 H), 1.78 (s, 3 H).

General procedure for preparation of Compound 5A

[0475] To a solution of Compound 4A (300 g, 771 mmol) in DCE (1.50 L) is added TMSOTf (257 g, 1.16 mol, 209 mL) and stirred for 2 hrs at 60 °C, and then stirred for 1 hr at 25 °C. Compound 2A (203 g, 848 mmol) is dissolved in DCE (1.50 L) and added 4 Å powder molecular sieves (150 g) stirring for 30 mins under N₂ atmosphere. Then the solution of Compound 4A in DCE is added dropwise to the mixture at 0 °C. The mixture is stirred at 25 °C for 16 hrs under N₂ atmosphere. TLC (DCM: MeOH = 25: 1, PMA) indicated Compound 4A is consumed completely and new spot (R_f = 0.24) formed. The reaction mixture is filtered and washed with sat. NaHCO₃ (2.00 L), water (2.00 L) and sat. brine (2.00 L). The organic layer is dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue is triturated with 2-Me-THE/heptane (5/3, v/v, 1.80 L) for 2 hrs, filtered and dried to give Compound 5A (225 g, 389 mmol, 50.3% yield, 98.4% purity) as a white solid. ¹H NMR: δ 7.81 (d, J = 9.29 Hz, 1 H), 7.20 - 7.42 (m, 6 H), 5.21 (d, J = 3.26 Hz, 1 H), 4.92 - 5.05 (m, 3 H), 4.55 (d, J = 8.28 Hz, 1 H), 3.98 - 4.07 (m, 3 H), 3.82 - 3.93 (m, 1 H), 3.71 - 3.81 (m, 1 H), 3.55 - 3.62 (m, 1 H), 3.43 - 3.53 (m, 2 H), 3.37 - 3.43 (m, 2 H), 3.14 (q, J = 5.77 Hz, 2 H), 2.10 (s, 3 H), 1.99 (s, 3 H), 1.89 (s, 3 H), 1.77 (s, 3 H).

General procedure for preparation of NAcegal-Linker-Tosylate salt

[0476] To a solution of Compound 5 A (200 g, 352 mmol) in THF (1.0 L) is added dry Pd/C (15.0 g, 10% purity) and TsOH (60.6 g, 352 mmol) under N₂ atmosphere. The suspension is degassed under vacuum and purged with H₂ several times. The mixture is stirred at 25 °C for 3 hrs under H₂ (45 psi) atmosphere. TLC (DCM: MeOH = 10: 1, PMA) indicated Compound 5A is consumed completely and one new spot (R_f = 0.04) is formed. The reaction mixture is filtered and concentrated (≤ 40 °C) under reduced pressure to give a residue. Diluted with anhydrous DCM (500 mL, dried overnight with 4 Å molecular sieves (dried at 300 °C for 12 hrs)) and concentrate to give a residue and run Karl Fisher (KF) to check for water content. This is repeated 3 times with anhydrous DCM (500 mL) dilutions and concentration to give NAcegal-Linker-TMSOTf (205 g, 95.8% yield, TsOH salt) as a foamy white solid. ¹ H NMR: δ 7.91 (d, J = 9.03 Hz, 1 H), 7.53 - 7.86 (m, 2 H), 7.49 (d, J = 8.03 Hz, 2 H),

7.13 (d, J = 8.03 Hz, 2 H), 5.22 (d, J = 3.26 Hz, 1 H), 4.98 (dd, J = 11.29, 3.26 Hz, 1 H), 4.57 (d, J =

8.53 Hz, 1 H), 3.99 - 4.05 (m, 3 H), 3.87 - 3.94 (m, 1 H), 3.79 - 3.85 (m, 1 H), 3.51 - 3.62 (m, 5 H), 2.96 (brt, J = 5.14 Hz, 2 H), 2.29 (s, 3 H), 2.10 (s, 3 H), 2.00 (s, 3 H), 1.89 (s, 3 H), 1.78 (s, 3 H). Scheme for the preparation of TRIS-PEG2-CBZ

General procedure for preparation of Compound 5B

[0477] To a solution of Compound 4B (400 g, 1.67 mol, 1.00 eq) and NaOH (10 M, 16.7 mL, 0.10 eq) in THF (2.00 L) is added Compound 4B_2 (1.07 kg, 8.36 mol, 1.20 L, 5.00 eq), the mixture is stirred at 30 °C for 2 hrs. LCMS showed the desired MS is given. Five batches of solution are combined to one batch, then the mixture is diluted with water (6.00 L), extracted with ethyl acetate (3.00 L*3), the combined organic layer is washed with brine (3.00 L), dried over Na₂SO₄, filtered and concentrated under vacuum. The crude is purified by column chromatography ( Si O₂. petroleum ether : ethyl acetate=l 00: 1-10: 1, R_f=0.5) to give Compound 5B (2.36 kg, 6.43 mol, 76.9% yield) as light yellow oil. HNMR: δ 7.31-7.36 (m, 5 H), 5.38 (s, 1 H), 5.11-5.16 (m, 2 H), 3.75 (t, J=6.4 Hz), 3.54- 3.62 (m, 6 H), 3.39 (d, J=5.2 Hz), 2.61 (t, J=6.0 Hz).

General procedure for preparation of 3-oxo-l-phenyl-2, 7, 10-trioxa-4-azatridecan- 13-oic acid (Compound 2B below)

[0478] To a solution of Compound 5B (741 g, 2.02 mol, 1.00 eq) in DCM (2.80 L) is added TFA (1.43 kg, 12.5 mol, 928 mL, 6.22 eq), the mixture is stirred at 25 °C for 3 hrs. LCMS showed the desired MS is given. The mixture is diluted with DCM (5.00 L), washed with water (3.00 L*3), brine (2.00 L), the combined organic layer is dried over Na₂SO₄, filtered and concentrated under vacuum to give Compound 2B (1800 g, crude) as light yellow oil. HNMR: δ 9.46 (s, 5 H), 7.27-7.34 (m, 5 H), 6.50-6.65 (m, 1 H), 5.71 (s, 1 H), 5.10-5.15 (m, 2 H), 3.68-3.70 (m, 14 H), 3.58-3.61 (m, 6 H), 3.39 (s, 2 H), 2.55 (s, 6 H), 2.44 (s, 2 H).

General procedure for preparation of Compound 3B

1B

[0479] To a solution of Compound 2B (375 g, 999 mmol, 83.0% purity, 1.00 eq) in DCM (1. 80 L) is added HATU (570 g, 1.50 mol, 1.50 eq) and DIEA (258 g, 2.00 mol, 348 mL, 2.00 eq) at 0 °C, the mixture is stirred at 0 °C for 30 min, then Compound IB (606 g, 1.20 mol, 1.20 eq) is added, the mixture is stirred at 25 °C for 1 hr. LCMS showed desired MS is given. The mixture is combined to one batch, then the mixture is diluted with DCM (5.00 L), washed with 1 N HCl aqueous solution (2.00 L*2), then the organic layer is washed with saturated Na₂CO₃ aqueous solution (2.00 L *2) and brine (2.00 L), the organic layer is dried over Na₂SO₄, filtered and concentrated under vacuum to give Compound 3B (3.88 kg, crude) as yellow oil.

General procedure for preparation of TRIS-PEG2-CBZ.

[0480] A solution of Compound 3B (775 g, 487 mmol, 50.3% purity, 1.00 eq) in HCl/dioxane (4 M, 2.91 L, 23.8 eq) is stirred at 25 °C for 2 hrs. LCMS showed the desired MS is given. The mixture is concentrated under vacuum to give a residue. Then the combined residue is diluted with DCM (5.00 L), adjusted to pH=8 with 2.5 M NaOH aqueous solution, and separated. The aqueous phase is extracted with DCM (3.00 L) again, then the aqueous solution is adjusted to pH=3 with 1 N HCl aqueous solution, then extracted with DCM (5.00 L*2), the combined organic layer is washed with brine (3.00 L), dried over Na₂SO₄, filtered and concentrated under vacuum. The crude is purified by column chromatography (SiO₂, DCM:MeOH=0: 1-12: 1, 0.1% HOAc, Rr=0.4). The residue is diluted with DCM (5.00 L), adjusted to pH=8 with 2.5 M NaOH aqueous solution, separated, the aqueous solution is extracted with DCM (3.00 L) again, then the aqueous solution is adjusted to pH=3 with 6 N HCl aqueous solution, extracted with DCM:MeOH=10: 1 (5.00 L*2), the combined organic layer is washed with brine (2.00 L), dried over Na₂SO₄, filtered and concentrated under vacuum to give a residue. Then the residue is diluted with MeCN (5.00 L), concentrated under vacuum, repeat this procedure twice to remove water to give TRIS-PEG2-CBZ (1.25 kg, 1.91 mol, 78.1% yield, 95.8% purity) as light yellow oil. ¹ HNMR: 400 MHz, MeOD, δ 7.30-7.35 (5 H), 5.07 (s, 2 H), 3.65-3.70 (m, 16 H), 3.59 (s, 4 H), 3.45 (t, J=5.6 Hz), 2.51 (t, J=6.0 Hz), 2.43 (t, 6.4 Hz).

Scheme for the preparation of TriNGal-TRIS-Peg2-Phosph 8c

TriGNal-TRIS-Peg2-Phosph 8c

General procedure for preparation of Compound 3C

[0481] To a solution of Compound 1C (155 g, 245 mmol, 1.00 eq) in ACN (1500 mL) is added

TBTU (260 g, 811 mmol, 3.30 eq), DIEA (209 g, 1.62mol, 282 mL, 6.60 eq) and Compound 2C (492 g, 811 mmol, 3.30 eq, TsOH) at 0 °C, the mixture is stirred at 15 °C for 16 hrs. LCMS showed the desired MS is given. The mixture is concentrated under vacuum to give a residue, then the mixture is diluted with DCM (2000 mL), washed with 1 N HCl aqueous solution (700 mL * 2), then saturated NaHCO₃ aqueous solution (700 mL *2) and concentrated under vacuum. The crude is purified by column chromatography to give Compound 3C (304 g, 155 mmol, 63.1% yield, 96.0% purity) as a yellow solid.

General procedure for preparation of Compound 4C

[0482] Two batches solution of Compound 3C (55.0 g, 29.2 mmol, 1.00 eq) in MeOH (1600 mL) is added Pd/C (6.60 g, 19.1 mmol, 10.0 % purity) and TFA (3.34 g, 29.2 mmol, 2.17 mL, 1.00 eq), the mixture is degassed under vacuum and purged with H₂. The mixture is stirred under H₂ (15 psi) at 15 °C for 2 hours. LCMS showed the desired MS is given. The mixture is filtered and the filtrate is concentrated under vacuum to give Compound 4C (106 g, 54.8 mmol, 93.7% yield, 96.2% purity, TFA) as a white solid.

General procedure for preparation of compound 5C

[0483] Two batches in parallel. To a solution of EDCI (28.8 g, 150 mmol, 1.00 eq) in DCM (125 mL) is added compound 4a (25.0 g, 150 mmol, 1.00 eq) dropwise at 0 °C, then the mixture is added to compound 4 (25.0 g, 150 mmol, 1.00 eq) in DCM (125 mL) at 0 °C, then the mixture is stirred at 25 °C for 1 hr. TLC (Petroleum ether : Ethyl acetate = 3 : 1, R_f = 0.45) showed the reactant is consumed and one new spot is formed. The reaction mixture is diluted with DCM (100 mL) then washed with aq.NaHCO₃ (250 mL * 1) and brine (250 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue is purified by column chromatography (SiO₂, Petroleum ether : Ethyl acetate = 100 : 1 to 3 : 1), TLC (SiCL, Petroleum ether : Ethyl acetate = 3: 1), R_f= 0.45, then concentrated under reduced pressure to give a residue.

Compound 5C (57.0 g, 176 mmol, 58.4% yield, 96.9% purity) is obtained as colorless oil and confirmed ¹ HNMR: EW33072-2-P1A, 400 MHz, DMSO δ 9.21 (s, 1 H), 7.07-7.09 (m, 2 H), 6.67-6.70 (m, 2 H), 3.02-3.04 (m, 2 H), 2.86-2.90 (m, 2 H).

General procedure for preparation of compound 6

[0484] To a mixture of compound 3 (79.0 g, 41.0 mmol, 96.4% purity, 1.00 eq, TFA) and compound 6C (14.2 g, 43.8 mmol, 96.9% purity, 1.07 eq) in DCM (800 mL) is added TEA (16.6 g, 164 mmol, 22.8 mL, 4.00 eq) dropwise at 0 °C, the mixture is stirred at 15 °C for 16 hrs. LCMS (EW33072-12- P IB, Rt = 0.844 min) showed the desired mass is detected. The reaction mixture is diluted with DCM (400 mL) and washed with aq.NaHCO₃ (400 mL * 1) and brine(400 mL * 1), then the mixture is diluted with DCM (2.00 L) and washed with 0.7 M Na₂CO₃ (1000 mL * 3) and brine(800 mL * 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue is used to next step directly without purification. Compound 6 (80.0 g, crude) is obtained as white solid and confirmed via ¹ HNMR: EW33072-12-P1A, 400 MHz, MeOD δ 7.02 - 7.04 (m, 2 H), 6.68 - 6.70 (m, 2 H), 5.34 - 5.35 (s, 3 H), 5.07 - 5.08 (d, J= 4.00 Hz, 3 H), 4.62 - 4.64 (d, J= 8.00 Hz, 3 H), 3.71 - 4. 16 (m, 16 H), 3.31 - 3.70 (m, 44 H), 2.80 - 2.83 (m, 2 H), 2.68 (m, 2 H), 2.46 - 2.47 (m, 10 H), 2.14 (s, 9 H), 2.03 (s, 9 H), 1.94 - 1.95 (d, J= 4.00 Hz, 18 H). General procedure for preparation of TriGNal-TRIS-Peg2-Phosph 8c

[0485] Two batches are synthesized in parallel. To a solution of compound 6C (40.0 g, 21.1 mmol,

1.00 eq in DCM (600 mL) is added diisopropylammonium tetrazolide (3.62 g, 21. 1 mmol, 1.00 eq) and compound 7c (6.37 g, 21. 1 mmol, 6.71 mL, 1.00 eq) in DCM (8.00 mL) drop-wise, the mixture is stirred at 30 °C for 1 hr, then added compound 7c (3.18 g, 10.6 mmol, 3.35 mL, 0.50 eq) in DCM

(8.00 mL) drop-wise, the mixture is stirred at 30 °C for 30 mins, then added compound 7c (3. 18 g, 10.6 mmol, 3.35 mL, 0.50 eq) in DCM (8.00 mL) drop-wise, the mixture is stirred at 30 °C for 1.5 hrs. LCMS (EW33072-17-P1C1, Rt = 0.921 min) showed the desired MS+1 is detected. LCMS (EW33072-17-P1C2, Rt = 0.919 min) showed the desired MS+1 is detected. Two batches are combined for work-up. The mixture is diluted with DCM (1.20 L), washed with saturated NaHCO₃ aqueous solution (1.60 L * 2), 3% DMF in H₂O (1.60 L * 2), H₂O (1.60 L * 3), brine (1.60 L), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue is purified by column chromatography (SiO₂, DCM : MeOH : TEA = 100 : 3 : 2) TLC (SiO₂, DCM: MeOH = 10: 1, R_f = 0.45), then concentrated under reduced pressure to give a residue. Compound 8C (76.0 g, 34.8 mmol, 82.5% yield, 96.0% purity) is obtained as white solid and confirmed via ¹ HNMR: EW33072-19-P1C, 400 MHz, MeOD δ 7. 13-7. 15 (d, J= 8.50 Hz, 2 H), 6.95-6.97 (dd, J =8.38, 1. 13 Hz, 2 H), 5.34 (d, J =2.88 Hz, 3 H),.09 (dd, J =11.26, 3.38 Hz, 3 H), 4.64 (d, J =8.50 Hz, 3 H), 3.99 - 4.20 (m, 12 H), 3.88 - 3.98 (m, 5 H), 3.66 - 3.83 (m, 20 H), 3.51 - 3.65 (m, 17 H), 3.33 - 3.50 (m, 9 H), 2.87 (t, J =7.63 Hz, 2 H), 2.76 (t, J =5.94 Hz, 2 H), 2.42 - 2.50 (m, 10 H), 2. 14 (s, 9 H), 2.03 (s, 9 H), 1.94 - 1.95 (d, J =6. 13 Hz, 18 H), 1.24-1.26 (d, J =6.75 Hz, 6 H), 1.18-1.20 (d, J =6.75 Hz, 6 H).

Example 11: Modification motif 1

[0486] An example GPAM siRNA includes a combination of the following modifications:

• Position 9 (from 5’ to 3’) of the sense strand is 2’ F.

• If position 9 is a pyrimidine then all purines in the Sense Strand are 2’OMe, and 1 -5 pyrimidines between positions 5 and 11 are 2’ F provided that there are never three 2’F modifications in a row.

• If position 9 is a purine then all pyrimidines in the Sense Strand are 2’OMe, and 1 -5 purines between positions 5 and 11 are 2’ F provided that there are never three 2’F modifications in a row.

• Antisense strand odd-numbered positions are 2'OMe and even-numbered positions are a mixture of 2’ F, 2’OMe and 2’ deoxy.

Example 12: Modification motif 2

[0487] An example GPAM siRNA includes a combination of the following modifications:

• Position 9 (from 5’ to 3’) of the sense strand is 2’ deoxy.

• Sense strand positions 5, 7 and 8 are 2’ F.

• All pyrimidines in positions 10-21 are 2’ OMe, and purines are a mixture of 2’ OMe and 2’ F. Alternatively, all purines in positions 10-21 are 2’ OMe and all pyrimidines in positions 10-21 are a mixture of 2’ OMe and 2’ F.

• Antisense strand odd-numbered positions are 2'OMe and even-numbered positions are a mixture of 2’ F, 2’OMe and 2’ deoxy. Example 13. Screening of siRNAs ETD01994-ETD02018 targeting human GPAM mRNA in mice transfected with AAV8-TBG-h-GPAM

[0488] The activities of siRNAs, namely ETD01994-ETD02018, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 13, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 14. ETD01994-ETD02004 were tested in Part 1 of the study and ETD02005-ETD02018 were tested in Part 2.

[0489] Six to eight week old female mice (C57B1/6) were inj ected with 5 μL of a recombinant adeno- associated virus 8 (AAV8) vector (1.3 x 10E13 genome copies/mL) by the retroorbital route on Day - 14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=4).

[0490] Mice were euthanized on Day 10 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml), or mouse GPAM (ThermoFisher, assay# Mm01261106_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_g I ). and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Only ETD01997, ETD01998, ETD02003, ETD02010, ETD02018 have highly similar target sequences in human and mouse GPAM. Data were normalized to the mean GPAM mRNA level in animals receiving a subcutaneous injection of PBS. Results for Part 1 are shown in Table 15, and those for Part 2 in Table 16. Of the siRNAs in this screening set, mice injected with ETD02009, ETD02011, ETD02012 or ETD02015 had the highest level of human GPAM mRNA knockdown in the liver. Table 13. Example siRNA Sequences

Table 14. Example siRNA Base Sequences

Table 15. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM - Part 1

Table 16. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM - Part 2

Example 14. Screening siRNAs with alternative modification patterns of ETD02012 and

ETD02011 in mice transfected with AAV8-TBG-h-GPAM

[0491] The base sequences of ETD02012 and ETD02011 were synthesized to generate siRNAs (ETD02012, ETD02285-ETD02292 and ETD02011, ETD02301-ETD02309, respectively) with alternative modification paterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 17, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 18.

[0492] Six to eight week old female mice (C57B1/6) were inj ected with 5 μL of a recombinant adeno- associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) by the retroorbital route on Day - 14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0493] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_g I ), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM, as defined by having a Ct value of >30, were omited from further analysis. Data were normalized to the mean GPAMmRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in Table 19. Of the alternatively modified versions of ETD02012, ETD02289 had the greatest activity. Of the alternatively modified versions of ETD02011, ETD02304 and ETD02309 had the greatest activity.

Table 17. Example siRNA Sequences

Table 18. Example siRNA Base Sequences

Table 19. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM

Example 15. Screening siRNAs with alternative modification patterns of ETD02009 and ETD02015 in mice transfected with AAV8-TBG-h-GPAM

[0494] The base sequences of ETD02009 and ETD02015 were synthesized to generate siRNAs (ETD02293-ETD02300, and ETD02015-ETD02318, respectively) with alternative modification patterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 20, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 21.

[0495] Six to eight week old female mice (C57B1/6) were inj ected with 5 μL of a recombinant adeno- associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) by the retroorbital route on Day - 14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0496] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_g I ), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM, as defined by having a Ct value of >30, were omitted from further analysis. Data were normalized to the mean GPAMmRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in Table 22. Of the alternatively modified versions of ETD02009, ETD02296 had the greatest activity. Of the alternatively modified versions of ETD02015, ETD02318 had the greatest activity. Table 20. Example siRNA Sequences

Table 21. Example siRNA Base Sequences

Table 22. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM

Example 16. Screening siRNAs with alternative modification patterns of ETD02003 and ETD02018 in mice transfected with AAV8-TBG-h-GPAM

[0497] The base sequences of ETD02003 and ETD02018 were synthesized to generate siRNAs (ETD02223-ETD02230, and ETD02201-ETD02208, respectively) with alternative modification patterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 23, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 24

[0498] Six to eight week old female mice (C57B1/6) were injected with 5 μL of a recombinant adeno- associated virus 8 (AAV8) vector (1.3 x 10E13 genome copies/mL) by the retroorbital route on Day - 14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single lOOμg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=4).

[0499] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) or mouse GPAM (ThermoFisher, assay# Mm01261106_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm()2342430_g I ). and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data were normalized to the mean GPAM mRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in Table 25. Of the alternatively modified versions of ETD02003, ETD02224 and ETD02228 had the greatest activity in terms of knockdown of the human GPAM mRNA. Of the alternatively modified versions of ETD02018, ETD02202 had the greatest activity in terms of knockdown of the human GPAM mRNA.

Table 23. Example siRNA Sequences

Table 24. Example siRNA Base Sequences

Table 25. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM

Example 17. Screening modification patterns of ETD02009 and ETD02015 containing 2’-O-(2- methoxyethyl) in mice transfected with AAV8-TBG-h-GPAM

[0500] The base sequences of ETD02009 and ETD02015 were synthesized to generate siRNAs (ETD02543-ETD02544, and ETD02539-ETD02542, respectively) with alternative modification patterns including 2’-O-(2-methoxyethyl). The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 26, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2- methoxyethyl) modified nucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 27.

[0501] Six to eight week old female mice (C57B1/6) were inj ected with 5 μL of a recombinant adeno- associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) by the retroorbital route on Day - 14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0502] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs00326039_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_g I ), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Data were normalized to the mean GPAM mRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in Table 28. Of the alternatively modified versions of ETD02015, ETD02541 had the greatest activity. Of the alternatively modified versions of ETD02009, ETD02543 and ETD2544 had the greatest activity. Table 26. Example siRNA Sequences

Table 27. Example siRNA Base Sequences

Table 28. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM

Example 18. Screening of siRNAs ETD02553-ETD02576 targeting human GPAM mRNA in mice transfected with AAV8-TBG-h-GPAM

[0503] The activities of siRNAs, namely ETD02553-ETD02576, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 29, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2- methoxyethyl) modified nucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 30. ETD02553-ETD02664 were tested in Part 1 of the study and ETD02565-ETD02576 were tested in Part 2.

[0504] Six to eight week old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (1.3 x l0E13 genome copies/mL) by the retroorbital route on Day -14. The recombinant AAV8 contains the open reading frame, the 5’ UTR, and a portion of the 3’UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0505] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_gl), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM in Part 2, as defined by having a Ct value of >32, were omitted from further analysis. Data were normalized to the mean GPAM mRNA level in animals receiving a subcutaneous injection of PBS. Results for Part 1 are shown in Table 31, and those for Part 2 in Table 32. Of the siRNAs in this screening set, mice injected with ETD02557, ETD02562, ETD02563, ETD02564, ETD02572, ETD02573 or ETD02574 had the highest level of human GPAM mRNA knockdown in the liver.

Table 29. Example siRNA Sequences

Table 30. Example siRNA Base Sequences

Table 31. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM - Part 1

Table 32. Relative GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB-h- GPAM - Part 2

Example 19: Therapeutic siRNA-mediated knockdown of GPAM in a mouse model of nonalcoholic fatty liver disease (NAFLD)

[0506] The effects of siRNA-mediated knockdown of GPAM in the liver was investigated in a high fat, high fructose diet-based mouse model of non-alcoholic fatty liver disease (NAFLD). Six- to eight- week-old C57BL/6NHsd female mice (Envigo) were given a Western high fat diet (Envigo TD. 120330, 0.2% cholesterol, 45% fat by calories) and high fructose water (55% fructose, 45% glucose) for eight weeks (Groups 2-4) or sixteen weeks (Groups 6-8) prior to siRNA injection. [0507] After 8 weeks, mice in Group 1 on regular chow (n=5) and Group 2 on Western Diet (n=5) were injected with 100 μL of phosphate buffered saline (PBS) subcutaneously biweekly until 16 weeks (Day 112). Mice in Group 3 on Western Diet (n=5) were injected with 100 μg of siRNA ETD02282 targeting mouse GPAM and Group 4 on Western Diet (n=5) were injected with 100 μg of siRNA ETD02284 targeting mouse GPAM by subcutaneous injection biweekly until 16 weeks (Day 112) for a total of 4 injections. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 33, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 34.

Table 33. Example siRNA Sequences

Table 34. Example siRNA Base Sequences

[0508] After 16 weeks, mice in Group 5 on regular chow (n=5) and Group 6 on Western Diet (n=5) were injected with 100 μL of phosphate buffered saline (PBS) subcutaneously biweekly until 24 weeks (Day 168). Mice in Group 7 on Western Diet (n=5) were injected with 100 μg of siRNA ETD02282 targeting mouse GPAM and Group 8 on Western Diet (n=5) were injected with 100 μg of siRNA ETD02284 targeting mouse GPAM by subcutaneous injection biweekly until 24 weeks (Day 168) for a total of 4 injections. The

[0509] Body weights were recorded biweekly at the start of the Western diet until the end of the study. Mice were fasted 4-6 hours for serum collection prior to the first siRNA treatment and then 1 day prior to each biweekly siRNA injection. Serum was sent for the following clinical chemistry assays (IDEXX Laboratories, Incorporated): ALP, ALT, BUN, cholesterol, glucose, total bilirubin, total protein, triglycerides, and betahydroxy butyrate. Ketones were measured in a drop of whole blood using Precision Xtra Ketone Test Strips (Abbott). [0510] All mice in Groups 1-4 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 16 weeks and all mice in Groups 5-8 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 24 weeks and the liver was separated into 4 sections. One section was placed in RNAlater for qRT-PCR (Thermo Fisher #AM7020), one section was snap frozen in liquid nitrogen for liver triglyceride measurement, one section was placed into 10% formalin for fixation and paraffin embedding for H&E and Picrosirius Red staining, and one section was fixed with 10% formalin at 4C for 24 hours and then embedded in OCT for Oil Red O staining.

[0511] Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’ s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048- 500) according to the manufacturer’ s instructions. The levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for mouse GPAM (ThermoFisher, assay# Mm01261106_ml), mouse COL1A1 (ThermoFisher, assay # Mm00801666_g I ). mouse TIMP1 (ThermoFisher, assay # Mm00801666_g I ), mouse TGFB1 (ThermoFisher, assay # Mm01341361_ml), mouse Acta2/SMA (ThermoFisher, assay # Mm00725412_sl), mouse Tnf-α (ThermoFisher, assay # Mm00443258_ml), mouse I11B (ThermoFisher, assay # Mm00434228_ml), mouse CCL2 (ThermoFisher, assay # Mm00441242_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm()2342430_g I ) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data was normalized to the mouse group on Western Diet receiving PBS (Group 2) by the delta-delta Ct method.

[0512] The 16 week GPAM mRNA levels are shown in Table 35. Data were normalized to the level in animals receiving PBS and on the Western Diet (Group 2). Mice on the Western Diet treated with ETD02282 (Group 3) or ETD02284 (Group 4) had reduced liver GPAM mRNA levels compared to mice on the Western Diet receiving PBS (Group 2). The mice on the Western Diet had elevated levels of GPAM compared to the mice on normal chow.

Table 35. 16-week GPAM mRNA Levels in Mice Treated with ETD02282 and ETD02284

[0513] The 24 -week GPAM mRNA levels are shown in Table 36. Data were normalized to the level in animals receiving PBS and on the Western Diet (Group 6). Mice on the Western Diet treated with ETD02282 (Group 7) or ETD02284 (Group 8) had reduced liver GPAM mRNA levels compared to mice on the Western Diet receiving PBS (Group 6). The mice on the Western Diet had elevated levels of GPAM compared to the mice on normal chow.

Table 36. 24-week GPAM mRNA Levels in Mice Treated with ETD02282 and ETD02284

[0514] The starting (Day 0) and ending (Day 112) body weights are shown in Table 37 for the 16- week study. There was reduced weight gain in the mice fed the Western Diet and treated with ETD02284 (Group 4) compared to mice on the Western Diet and treated with PBS (Group 2) at 16 weeks. Table 37. 16-week Body Weights of Mice Treated with ETD02282 and ETD02284

[0515] The starting (Day 0) and ending (Day 168) body weights are shown in Table 38 for the 24- week study.

Table 38. 24-week Body Weights of Mice Treated with ETD02282 and ETD02284

[0516] The 16-week serum levels of ALT, cholesterol, and ketones are shown in Table 39. The ALT and cholesterol levels at 16 weeks in mice treated with ETD02282 (Group 3) and ETD02284 (Group 4) were lower than mice treated with PBS (Group 2) on Western Diet. The ketone levels in mice at 16 weeks treated with ETD02282 (Group 3) and ETD02284 (Group 4) were higher than mice treated with PBS (Group 2) on Western Diet.

Table 39. 16-week ALT, Cholesterol, and Ketone Levels in Mice Treated with ETD02282 and ETD02284

[0517] The 24-week serum levels of ALT, cholesterol, and ketones are shown in Table 40. The ALT and cholesterol levels at 24 weeks in mice treated with ETD02282 (Group 7) and ETD02284 (Group 8) were lower than mice treated with PBS (Group 6) on Western Diet. The ketone levels in mice at 16 weeks treated with ETD02282 (Group 7) and ETD02284 (Group 8) were higher than mice treated with PBS (Group 6) on Western Diet.

Table 40. 24-week ALT, Cholesterol, and Ketone Levels in Mice Treated with ETD02282 and ETD02284

[0518] To assess liver histology, liver sections were formalin fixed and paraffin embedded then stained with hematoxylin and eosin (H&E). The % steatotic area was calculated by measuring the white areas left behind by lipid droplets in the tissue divided by the total area of the tissue section. Five different liver serial sections were used per animal for quantification. The 16-week % steatotic area of the H&E stained formalin fixed liver sections is in Table 41. The % steatotic area at 16 weeks was reduced in mice treated with ETD02282 (Group 3) and ETD02284 (Group 4) compared to mice treated with PBS (Group 2) on Western Diet.

Table 41. 16-week % Steatotic Area of Livers in Mice Treated with ETD02282 and ETD02284

[0519] To assess liver histology, liver sections were formalin fixed and paraffin embedded then stained with hematoxylin and eosin (H&E). The % steatotic area was calculated by measuring the white areas left behind by lipid droplets in the tissue divided by the total area of the tissue section. Five different liver serial sections were used per animal for quantification. The 24-week % steatotic area of the H&E stained formalin fixed liver sections is in Table 42. The % steatotic area at 24 weeks was reduced in mice treated with ETD02282 (Group 7) and ETD02284 (Group 8) compared to mice treated with PBS (Group 6) on Western Diet. Table 42. 24-week % Steatotic Area of Livers in Mice Treated with ETD02282 and ETD02284

[0520] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride Colorimetric Kit (Elabscience #E-BC-K238) following the manufacturer’s instructions. The relative levels of triglycerides in the flash frozen liver sections are shown in Table 43 with the data normalized to mice on normal diet treated with PBS (Group 1). The level of triglycerides in the liver at 16 weeks was reduced in mice treated with ETD02282 (Group 3) and ETD02284 (Group 4) compared to mice treated with PBS (Group 2) on Western Diet.

Table 43. 16-week Liver Triglyceride Levels in Mice Treated with ETD02282 and ETD02284

[0521] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride

Colorimetric Kit (Elabscience #E-BC-K238) following the manufacturer’s instructions. The levels of triglycerides in the flash frozen liver sections are shown in Table 44 with the data normalized to mice on normal diet treated with PBS (Group 5). The level of triglycerides in the liver at 24 weeks was reduced in mice treated with ETD02284 (Group 8) compared to mice treated with PBS (Group 6) on Western Diet.

Table 44. 24-week Liver Triglyceride Levels in Mice Treated with ETD02282 and ETD02284

[0522] Expression of fibrotic and pro-inflammatory genes in the liver were measured by qRT-PCR and normalized to the mice on Western Diet receiving PBS (Group 2). The expression of pro-fibrotic genes COL1A1 and TIMP1 and pro-inflammatory genes TNF-α, IL1b, and CCL2 are shown in Table 45. Mice on Western Diet treated with ETD02284 (Group 4) had reduced expression of pro-fibrotic genes COL1A1 and TIMP1 and pro-inflammatory genes TNF-alpha and CCL2 at 16 weeks compared to mice treated with PBS (Group 2) on Western Diet.

Table 45. 16-week Expression of Pro-fibrotic and Pro-inflammatory Genes in Mice Treated with ETD02282 and ETD02284

[0523] Expression of fibrotic and pro-inflammatory genes in the liver were measured by qRT-PCR and normalized to the mice on Western Diet receiving PBS (Group 2). The expression of pro -fibrotic genes COL1A1 and TIMP1 and pro-inflammatory genes TNF-α, IL1b, and CCL2 are shown in Table 46. Mice on Western Diet treated with ETD02282 (Group 7) and ETD02284 (Group 8) had reduced expression of pro-fibrotic genes COL1A1 and TIMP1 and pro-inflammatory gene TNF-alpha at 24 weeks compared to mice treated with PBS (Group 6).

Table 46. 24-week Expression of Pro-fibrotic and Pro-inflammatory Genes in Mice Treated with ETD02282 and ETD02284

Example 20: Preventative siRNA-mediated knockdown of GPAM in a mouse model of nonalcoholic fatty liver disease (NAFLD)

[0524] The protective effects of siRNA-mediated knockdown of GPAM in the liver was investigated in a high fat, high fructose diet-based mouse model of non-alcoholic fatty liver disease (NAFLD).

Six- to eight- week-old C57BL/6NHsd female mice (Envigo) were given a Western high fat diet (Envigo TD. 120330, 0.2% cholesterol, 45% fat by calories) and high fructose water (55% fructose, 45% glucose). Mice were on diet for eight weeks (Groups 2-6) or sixteen weeks (Groups 8-12) with the following siRNAs targeting GPAM starting Day 0 of the study: Group 1 (n=5) on regular chow were injected with 100 μL of phosphate buffered saline (PBS) subcutaneously biweekly until 8 weeks (Day 56) or Group 7 (n=5) until 16 weeks (Day 112). Group 2 (n=5) on the Western diet were injected with 100 μL of phosphate buffered saline (PBS) subcutaneously biweekly until 8 weeks (Day 56) or Group 8 (n=5) until 16 weeks (Day 112).

[0525] Group 3 (n=5) on the Western diet were injected with 200 μg of siRNA ETD02282 targeting mouse GPAM subcutaneously biweekly until 8 weeks (Day 56) or Group 9 (n=5) until 16 weeks (Day 112). Group 4 (n=5) on the Western diet were injected with 60 μg of siRNA ETD02282 targeting mouse GPAM subcutaneously biweekly until 8 weeks (Day 56) or Group 10 (n=5) until 16 weeks (Day 112). Group 5 (n=5) on the Western diet were injected with 200 μg of siRNA ETD02284 targeting mouse GPAM subcutaneously biweekly until 8 weeks (Day 56) or Group 11 (n=5) until 16 weeks (Day 112). Group 6 (n=5) on the Western diet were injected with 60 μg of siRNA ETD02284 targeting mouse GPAM subcutaneously biweekly until 8 weeks (Day 56) or Group 12 (n=5) until 16 weeks (Day 112). Mice on the 8 week study received a total of 4 siRNA doses and mice on the 16 week study received a total of 16 siRNA doses. The siRNA sequences that were used are shown in Table 47, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro- modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 48.

Table 47. Example siRNA Sequences

Table 48. Example siRNA Base Sequences

[0526] Body weights were recorded biweekly until the end of the study. Mice were fasted 4-6 hours for serum collection on Day 0 and then 1 day prior to each biweekly siRNA injection. Serum was sent for the following clinical chemistry assays performed at IDEXX Laboratories, Incorporated - ALP, ALT, BUN, cholesterol, glucose, total bilirubin, total protein, triglycerides, and beta-hydroxybutyrate. [0527] All mice in Groups 1-6 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 8 weeks and all mice in Groups 7-12 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 16 weeks and the liver was separated into 4 sections. One section was placed in RNAlater for qRT-PCR (Thermo Fisher #AM7020), one section was snap frozen in liquid nitrogen for liver triglyceride measurement, one section was placed into 10% formalin for fixation and paraffin embedding for H&E and Picrosirius Red staining, and one section was fixed with 10% formalin at 4C for 24 hours and then embedded in OCT for Oil Red O staining.

[0528] Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’ s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048- 500) according to the manufacturer’ s instructions. The levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for mouse GPAM (ThermoFisher, assay# Mm01261106_ml), mouse COL1A1 (ThermoFisher, assay # Mm00801666_g I ). mouse TIMP1 (ThermoFisher, assay # Mm00801666_g I ), mouse TGFB1 (ThermoFisher, assay # Mm01341361_ml), mouse Acta2/SMA (ThermoFisher, assay # Mm00725412_sl), mouse Tnf-α (ThermoFisher, assay # Mm00443258_ml), mouse II IB (ThermoFisher, assay # Mm00434228_ml), mouse CCL2 (ThermoFisher, assay # Mm00441242_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm()2342430_g 1 ) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data was normalized to the mouse group on Western Diet receiving PBS (Group 2) using the deltadelta Ct method.

[0529] The 8 -week GPAM mRNA levels are shown in Table 49. Data were normalized to the level in animals receiving PBS and on the Western Diet (Group 2). Mice on the Western Diet treated with ETD02282 (Groups 3 and 4) or ETD02284 (Groups 5 and 6) had reduced liver GPAM mRNA levels compared to mice on the Western Diet receiving PBS (Group 2). The mice on the Western Diet had elevated levels of GPAM compared to the mice on normal chow.

Table 49. 8-week GPAM mRNA Levels in Mice Treated with ETD02282 and ETD02284

[0530] The 16-week GPAM mRNA levels are shown in Table 50. Data were normalized to the level in animals receiving PBS and on the Western Diet (Group 8). Mice on the Western Diet treated with ETD02282 (Groups 9 and 10) or ETD02284 (Groups 11 and 12) had reduced liver GPAM mRNA levels compared to mice on the Western Diet receiving PBS (Group 8). The mice on the Western Diet had elevated levels of GPAM compared to the mice on normal chow.

Table 50. 16-week GPAM mRNA Levels in Mice Treated with ETD02282 and ETD02284

[0531] The starting (Day 0) and ending (Day 56) body weights are shown in Table 51 for the 8-week study. There was reduced weight gain in the mice fed the Western Diet and treated with both doses of ETD02284 (Groups 5 and 6) compared to mice on the Western Diet and treated with PBS (Group 2) at 8 weeks.

Table 51. 8-week Body Weights of Mice Treated with ETD02282 and ETD02284

[0532] The starting (Day 0) and ending (Day 112) body weights are shown in Table 52 for the 16- week study. There was reduced weight gain in mice fed the Western Diet and treated with 60 μg dose of ETD02282 (Group 10) and ETD02284 (Group 12) compared to mice on the Western Diet and treated with PBS (Group 8) at 16 weeks.

Table 52. 16-week Body Weights of Mice Treated with ETD02282 and ETD02284

[0533] The 8-week serum levels of ALT, cholesterol, and ketones are shown in Table 53. The ALT levels at 8 weeks in mice treated with both doses of ETD02282 (Groups 3 and 4) and ETD02284 (Groups 5 and 6) were lower than mice treated with PBS (Group 2) on Western Diet. The ketone levels in mice at 8 weeks treated with both doses of ETD02284 (Groups 5 and 6) on Western Diet were higher than mice treated with PBS (Group 2) on Western Diet.

Table 53. 8-week ALT, Cholesterol, and Ketone Levels in Mice Treated with ETD02282 and ETD02284

[0534] The 16-week serum levels of ALT, cholesterol, and ketone betahydroxybutyrate (BHB) are shown in Table 54. The ALT levels at 16 weeks in mice treated with both doses of ETD02282 (Groups 9 and 10) and ETD02284 (Groups 11 and 12) were lower than mice treated with PBS (Group 8) on Western Diet.

Table 54. 16-week ALT, Cholesterol, and Ketone Levels in Mice Treated with ETD02282 and ETD02284

[0535] To assess liver histology, liver sections were formalin fixed and paraffin embedded then stained with hematoxylin and eosin (H&E). The % steatotic area was calculated by measuring the white areas left behind by lipid droplets in the tissue divided by the total area of the tissue section. Five different liver serial sections were used per animal for quantification. The 8 -week % steatotic area of the H&E stained formalin fixed liver sections is in Table 55. Mice on the Western diet treated with the 200 μg dose of ETD02282 (Group 3) and both doses of ETD02284 (Groups 5 and 6) had reduced % steatotic area compared to mice received PBS (Group 2) on Western Diet.

Table 55. 8-week % Steatotic Area of Livers in Mice Treated with ETD02282 and ETD02284

[0536] To assess liver histology, liver sections were formalin fixed and paraffin embedded then stained with hematoxylin and eosin (H&E). The % steatotic area was calculated by measuring the white areas left behind by lipid droplets in the tissue divided by the total area of the tissue section. Five different liver serial sections were used per animal for quantification. The 16-week % steatotic area of the H&E stained formalin fixed liver sections is in Table 56. Mice on the Western Diet treated with both doses of ETD02282 (Groups 9 and 10) and ETD02284 (Groups 11 and 12) for 16 weeks had reduced % steatotic area compared to mice on Western Diet treated with PBS.

Table 56. 16-week % Steatotic Area of Livers in Mice Treated with ETD02282 and ETD02284

[0537] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride Colorimetric Kit (Elabscience #E-BC-K238) following the manufacturer’s instructions. The levels of triglycerides in the flash frozen liver sections are shown in Table 57 with the data normalized to mice on normal diet treated with PBS (Group 1). The level of triglycerides in the liver at 8 weeks was reduced in mice treated with 200 μg (Group 3) and 60 μg (Group 4) of ETD02282 and 200 μg (Group 5) and 60 μg (Group 6) of ETD02284 (Group 4) compared to mice treated with PBS (Group 2) on Western Diet.

Table 57. 8-week Liver Triglyceride Levels in Mice Treated with ETD02282 and ETD02284

[0538] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride Colorimetric Kit (Elabscience #E-BC-K238) following the manufacturer’s instructions. The levels of triglycerides in the flash frozen liver sections are shown in Table 58 with the data normalized to mice on normal diet treated with PBS (Group 7). The samples from the 60 μg dose of ETD02284 (Group 12) could not be successfully processed for this experiment.

Table 58. 16-week Liver Triglyceride Levels in Mice Treated with ETD02282 and ETD02284

[0539] Expression of fibrotic and pro-inflammatory genes in the liver were measured by qRT-PCR and normalized to the mice on Western Diet receiving PBS (Group 2). The expression of pro -fibrotic genes COL1A1 and TIMP1 and pro-inflammatory genes TNF-α, IL1b, and CCL2 are shown in Table 59. Mice on Western Diet treated with 60 μg of ETD02282 (Group 4) had reduced expression of pro- fibrotic genes COL1A1 and TIMP1 and reduced expression of CCL2 compared to mice treated with PBS (Group 2). Mice on Western Diet treated with 200 μg of ETD02284 (Group 5) and 60 μg (Group 6) had reduced expression of pro-fibrotic genes COL1A1 and TIMP1 and reduced expression of pro- inflammatory genes TNF-alpha, IL1b, and CCL2 when compared to mice treated with PBS (Group 2).

Table 59. 8-week Expression of Pro-fibrotic and Pro-inflammatory Genes in Mice Treated with ETD02282 and ETD02284

[0540] Expression of fibrotic and pro-inflammatory genes in the liver were measured by qRT-PCR and normalized to the mice on Western Diet receiving PBS (Group 2). The expression of pro -fibrotic genes C0L1A1 and TIMP1 and pro-inflammatory genes TNF-α, IL1b, and CCL2 are shown in Table 60. The mice treated with 60 μg of ETD02284 (Group 12) showed significant reduction of all pro- fibrotic and pro -inflammatory genes measured at 16 weeks compared to mice on PBS (Group 8).

Table 60. 16-week Expression of Pro-fibrotic and Pro-inflammatory Genes in Mice Treated with ETD02282 and ETD02284

Example 21: Determining the activity of a siRNA ETD02318 targeting GPAM in a single dose study in non-human primates

[0541] One group of four male cynomolgus monkeys >3 years old was utilized for this study. Monkeys were maintained on normal chow with ad libitum access throughout the study except prior to blood collections in which they were fasted overnight (at least 12 hours). On study Day 0, cynomolgus monkeys were injected subcutaneously (2 mL/kg) with a single dose of 3 mg/kg ETD02318 at a concentration of 1.5 mg/mL. The sequence of the siRNA used are shown in Table 61, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’- O-methyl modified nucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 62. Table 61. Example siRNA Sequence

Table 62. Example siRNA BASE Sequence

[0542] Body weights were recorded weekly on Days -8, -2, 7, 14, 21, and 28 of the study. On study Days -8, -2, 7, 14, 21, and 28 whole blood was collected into tubes with no anti -coagulant and centrifuged to obtain serum after clotting. Clinical chemistry for ALT, AST, ALP, DBIL, TBIL, GLU, UREA, CREA, TG, CHOL, TP, GGT, HDL-CH, LDL-CH, and B-HDBH were analyzed.

[0543] On study Day -8 and Day 28, a 5 mg liver biopsy was conducted by anesthetizing the animals with Zoletil (1.5-5.0 mg/kg, i.m.) andxylazine (0.5-2.0 mg/kg, i.m.). The liver biopsy was then placed into 10 v/v RNAlater™ Stabilization Solution (Thermo Fisher, Catalog# AM7020) in 20 seconds and stored for 24 hours at 4°C. The RNAlater was then removed and the liver tissue was stored in the freezer until analysis.

[0544] Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’ s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048- 500) according to the manufacturer’ s instructions. The levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for monkey GPAM (ThermoFisher, assay# Mf02878271_ml), and the monkey housekeeping gene GAPDH (ThermoFisher, assay# Mf()4392546_g I ) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222).

[0545] The results of the liver biopsy mRNA analysis are summarized in Table 63. Data for each individual was normalized to its Day -8 liver biopsy mRNA levels using the delta-delta Ct method. A 64% mean reduction in liver GPAM mRNA was observed with a single dose of ETD02318 at Day 28 after injection.

[0546] There were no significant changes in body weight or any of the clinical parameters measured during this study. Table 63. GPAM liver mRNA Levels in Monkeys treated with a single dose of ETD02318

Example 22: Bioinformatic selection of human GPAM siRNA sequences that are cross-reactive with different non-human primate species

[0547] Therapeutic siRNAs were designed to target human GPAM and, in some cases, the GPAM sequence of at least one toxicologically-relevant non-human primate species including cynomolgus monkey, marmoset, green monkey, or rhesus monkey. The siRNAs included in subset K have the following characteristics and are shown in Table 64:

• Cross-reactivity: With 19mer in human GPAM mRNA, with 17mer (pos. 2-18) in cynomolgus monkey, marmoset, green monkey, or rhesus monkey GPAM mRNA.

• SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18)

• No seed region matches to miRNAs conserved in human, mouse, and rat and not present in greater than 4 species.

Table 64: Subset K siRNAs

Example 23: Bioinformatic selection of additional human GPAM siRNA sequences that are cross-reactive with cynomolgus monkey GPAM

[0548] Therapeutic siRNAs were designed to target human GPAM having characteristics as described for Subset A, and the GPAM sequence of cynomolgus monkey. The siRNAs included in subset L have the following characteristics and are shown in Table 65.

Table 65: Subset L siRNAs

Example 24; Bioinformatic selection of human GPAM siRNA sequences that have one mismatch with cynomolgus monkey

[0549] Therapeutic siRNAs were designed to target human GPAM as described above for Subset A, and, in some cases, the GPAM sequence of cynomolgus monkey. Included in this set are siRNAs that have one mismatch with cynomolgus monkey GPAM. The siRNAs included in subset M have the following characteristics and are shown in Table 66.

Table 66: Subset M siRNAs

Example 25; Bioinformatic selection of additional human GPAM siRNA sequences that have one mismatch with cynomolgus monkey

[0550] Therapeutic siRNAs were designed to target human GPAM as described above for Subset A, and, in some cases, the GPAM sequence of cynomolgus monkey. Included in this set are siRNAs that have one mismatch with cynomolgus monkey GPAM. The siRNAs included in subset N have the following characteristics and are shown in Table 67.

Table 67: Subset N siRNAs

Example 26: Bioinformatic selection of human GPAM siRNA sequences that are cross- reactive with cynomolgus monkeys

[0551] Therapeutic siRNAs were designed to target human GPAM and, in some cases, and, in some cases, the GPAM sequence of cynomolgus monkey. The siRNAs included in subset O have the following characteristics and are shown in Table 68:

• Cross-reactivity: With 19mer in human GPAM mRNA, with 17mer (pos. 2-18) in cynomolgus monkey GPAM mRNA (Ensembl Transcript ID: ENSMFAT00000069191.2).

• Specificity: For human off-target transcripts expressed in hepatocytes AS2 or better and SS3 or better. AS2 or better and SS3 or better in cynomolgus monkey.

• SNPs: siRNA target sites do not harbor SNPs with a MAF ≥ 1% (pos. 2-18)

Table 68: Subset O siRNAs

[0552] In some cases, the antisense strand of any of the siRNAs of subset K comprise a modification or modification pattern. Some such examples are included in Table 69 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a,c,g,t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or uM) is a 2’-O-(2-methoxyethyl) modified nucleoside, “d” is a 2’ deoxynucleoside, “i” is a 2’-O-methyl inosine nucleoside, [NUNA] (e.g., AUNA, CUNA, GUNA, TUNA, or UUNA) is the unlocked nucleic acid version of the nucleoside, and “s” is a phosphorothioate linkage. The siRNAs in subset K may comprise any other modification pattem(s).

Table 69. Subset K siRNAs with Chemical Modifications

[0553] In some cases, the antisense strand of any of the siRNAs of subset L comprise a modification or modification pattern. Some such examples are included in Table 70 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a,c,g,t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, “d” is a 2’ deoxynucleoside, “i” is a 2’-O-methyl inosine nucleoside, [NUNA] (e.g., AUNA, CUNA, GUNA, TUNA, or UUNA) is the unlocked nucleic acid version of the nucleoside, and “s” is a phosphor othioate linkage. The siRNAs in subset L may comprise any other modification pattern(s).

Table 70. Subset L siRNAs with Chemical Modifications

[0554] In some cases, the antisense strand of any of the siRNAs of subset M comprise a modification or modification pattern. Some such examples are included in Table 71 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a,c,g,t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, “d” is a 2’ deoxynucleoside, “i” is a 2’-O-methyl inosine nucleoside, [NUNA] (e.g., AUNA, CUNA, GUNA, TUNA, or UUNA) is the unlocked nucleic acid version of the nucleoside, and “s” is a phosphor othioate linkage. The siRNAs in subset M may comprise any other modification pattem(s).

Table 71. Subset M siRNAs with Chemical Modifications

[0555] In some cases, the antisense strand of any of the siRNAs of subset N comprise a modification or modification pattern. Some such examples are included in Table 72 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a,c,g,t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, “d” is a 2’ deoxynucleoside, “i” is a 2’-O-methyl inosine nucleoside, [NUNA] (e.g., AUNA, CUNA, GUNA, TUNA, or UUNA) is the unlocked nucleic acid version of the nucleoside, and “s” is a phosphorothioate linkage. The siRNAs in subset N may comprise any other modification pattem(s).

Table 72. Subset N siRNAs with Chemical Modifications

[0556] In some cases, the antisense strand of any of the siRNAs of subset O comprise a modification or modification pattern. Some such examples are included in Table 73 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g., a,c,g,t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, “d” is a 2’ deoxynucleoside, “i” is a 2’-O-methyl inosine nucleoside, [NUNA] (e.g., AUNA, CUNA, GUNA, TUNA, or UUNA) is the unlocked nucleic acid version of the nucleoside, and “s” is a phosphorothioate linkage. The siRNAs in subset O may comprise any other modification pattem(s).

Table 73. Subset O siRNAs with Chemical Modifications

Example 27. Screening siRNAs with alternative modification patterns of ETD02562 in mice transfected with AAV8-TBG-h-GPAM

[0557] The base sequences of ETD02562 were synthesized to generate siRNAs (ETD02562, ETD02728-ETD02731, and ETD02740-ETD02747) with alternative modification patterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 74, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 75. [0558] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) and 20 μL of PBS + 5% glycerol for a total 30 μL injection volume by the retroorbital route on Day -14. The recombinant AAV8 contains the open reading frame, the 5’ UTR, and a portion of the 3’UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single lOOμg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0559] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_gl), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM, as defined by having a Ct value of >30, were omitted from further analysis. Data were normalized to the mean GPAMmRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in for the anti-sense strand modifications in Table 76 and for the sense strand modifications in Table 77. Of the alternatively modified versions of ETD02562, ETD02729, ETD02742, and ETD02746 had the greatest activity. Table 74. Example siRNA Sequences

Table 75. Example siRNA BASE Sequences

Table 76. Relative human GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB- h-GPAM

Table 77. Relative human GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB- h-GPAM

Example 28: Screening siRNAs with alternative modification patterns of ETD02563 in mice transfected with AAV8-TBG-h-GPAM

[0560] The base sequences of ETD02563 were synthesized to generate siRNAs (ETD02563, ETD02732-ETD02735, and ETD02748-ETD02755) with alternative modification patterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 78, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 79. [0561] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) and 20 μL of PBS + 5% glycerol for a total 30 μL injection volume by the retroorbital route on Day -14. The recombinant AAV8 contains the open reading frame, the 5’ UTR, and a portion of the 3’UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single lOOμg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n=5).

[0562] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_gl), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM, as defined by having a Ct value of >30, were omitted from further analysis. Data were normalized to the mean GPAMmRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in for the anti-sense strand modifications in Table 80 and for the sense strand modifications in Table 81. Of the alternatively modified versions of ETD02563, ETD02735 had the greatest activity.

Table 78. Example siRNA Sequences

Table 79. Example siRNA BASE Sequences

Table 80. Relative human GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB- h-GPAM

Table 81. Relative human GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB- h-GPAM

Example 29: Screening siRNAs with alternative modification patterns of ETD02564 in mice transfected with AAV8-TBG-h-GPAM

[0563] The base sequences of ETD02564 were synthesized to generate siRNAs (ETD02564, ETD02736-ETD02739) with alternative modification patterns. The activities of the siRNAs were assessed using mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 82, where Nf (e. g. , Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoromodified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’ -O-(2-methoxy ethyl) modified nucleoside, dN (e. g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 83.

[0564] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (1.2 x 10E13 genome copies/mL) and 20 μL of PBS + 5% glycerol for a total 30 μL injection volume by the retroorbital route on Day -14. The recombinant AAV 8 contains the open reading frame, the 5 ’ UTR, and a portion of the 3 ’ UTR of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-GPAM). On Day 0, infected mice were given a subcutaneous injection of a single lOOμg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n=5).

[0565] Mice were euthanized on Day 14 after subcutaneous injection and a liver sample from each was collected and placed in RNAlater (ThermoFisher Catalog# AM7020) until processing. Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_gl), and PerfeCTa® qPCRFastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Mice that gave low liver expression of human GPAM, as defined by having a Ct value of >30, were omitted from further analysis. Data were normalized to the mean GPAMmRNA level in animals receiving a subcutaneous injection of PBS. Results are shown in in Table 84. Of the alternatively modified versions of ETD02564, ETD02739 had the greatest activity. Table 82. Example siRNA Sequences

Table 83. Example siRNA BASE Sequences

Table 84. Relative human GPAM mRNA Levels in Livers of Mice Transfected with AAV8-TGB- h-GPAM

Example 30: Screening human and different non-human primate siRNAs in vitro in Hep3B cells, primary cynomolgus hepatocytes, and primary human hepatocytes.

[0566] Therapeutic siRNAs were designed to target human GPAM and, in some cases, the GPAM sequence of at least one toxicologically-relevant non-human primate species including cynomolgus monkey, marmoset, green monkey, or rhesus monkey. The activities of the siRNAs were assessed in Hep3B cells using transfection reagent or in human or cynomolgus primary hepatocytes using free uptake through the ASGPR. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 85, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2- methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 86.

[0567] Hep3B (ATCC, HB-8064) cells in EMEM + 10% FBS (Cytiva) were plated 10,000 cells per well in a 96- well plate and grown to -70% confluence. Transfection complexes were made by mixing 10 nM of siRNA in 25 μL of Opti-MEM media (Thermo Scientific) and 1.5 μL of Lipofectamine RNAi-MAX (Fisher, 13778150) in 25 μL of Opti-MEM media together and incubating 5 minutes at room temperature. The siRNAs were prepared in triplicate and 10 μL of the transfection complexes was added to the cells. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828).

[0568] Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (Thermo Fisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 87. Of the siRNAs screened in Hep3Bs, most had some activity with transfection reagent and the best sequences were ETD02976, ETD02979, ETD02983, ETD02991, and ETD02993.

[0569] Primary human hepatocytes (BioIVT) were plated 50,000 cells per well in a 96 -well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty-four hours post thawing, hepatocytes were treated with lOul siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 88. Of the siRNAs screened in primary human hepatocytes, the best sequences were ETD02979, ETD02983, and ETD02993.

[0570] Primary cyno hepatocytes (BioIVT) were plated 50,000 cells per well in a 96-well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty -four hours post thawing, hepatocytes were treated with 10 μL siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for cynomolgus monkey GPAM (ThermoFisher, assay# Mf02878271_ml) and cynomolgus monkey housekeeping gene GUSB (ThermoFisher, assay# Mf04392669_ml) and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222)). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 89. Of the siRNAs screened in primary cyno hepatocytes, the best sequences were ETD02983, ETD02993, and ETD02996.

Table 85. Example siRNA Sequences

Table 86. Example siRNA Base Sequences

Table 87. Relative GPAM mRNA Levels in Hep3B cells

Table 88. Relative GPAM mRNA Levels in Primary Human Hepatocytes

Table 89. Relative GPAM mRNA Levels in Primary Cyno Hepatocytes

Example 31: Screening human and cyno (1 mismatch) siRNAs in vitro in Hep3B cells, primary cyno hepatocytes, and primary human hepatocytes.

[0571] Therapeutic siRNAs were designed to target human GPAM and cynomolgus monkey with one mismatch with cynomolgus monkey GPAM sequence. The activities of the siRNAs were assessed in Hep3B cells using transfection reagent or in human or cyno primary hepatocytes using free uptake through the ASGPR. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 90, where Nf (e.g., Af, Cf, Gf, Tf, orUf) is a 2’-fhroro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 91.

[0572] Hep3B (ATC, HB-8064) cells in EMEM + 10% FBS (Cytiva) were plated 10,000 cells per well in a 96- well plate and grown to -70% confluence. Transfection complexes were made by mixing 10 nM of siRNA in 25 μL of Opti-MEM media (Thermo Scientific) and 1.5 μL of Lipofectamine RNA-MAX (Fisher, 13778150) in 25 μL of Opti-MEM media together and incubating 5 minutes at room temperature. The siRNAs were prepared in triplicate and 10 μL of the transfection complexes was added to the cells. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828).

[0573] Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (Thermo Fisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 92. Of the siRNAs screened in Hep3Bs, most had some activity with transfection reagent and the best sequences were ETD03008, ETD03011, and ETD03013.

[0574] Primary human hepatocytes (BioIVT) were plated 50,000 cells per well in a 96 -well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty-four hours post thawing, hepatocytes were treated with 10ul siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 93. Of the siRNAs screened in primary human hepatocytes, the best sequences were ETD03011 and ETD03013.

[0575] Primary cyno hepatocytes (BioIVT) were plated 50,000 cells per well in a 96-well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty-four hours post thawing, hepatocytes were treated with 10 μL siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for cynomolgus monkey GPAM (ThermoFisher, assay# Mf02878271_ml) and cynomolgus monkey housekeeping gene GUSB (ThermoFisher, assay# Mf04392669_ml) and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222)). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 94. Of the siRNAs screened in primary cyno hepatocytes, the best sequences were ETD03011 and ETD03013.

Table 90. Example siRNA Sequences

Table 91. Example siRNA Base Sequences

Table 92. Relative GPAM mRNA Levels in Hep3B cells

Table 93. Relative GPAM mRNA Levels in Primary Human Hepatocytes

Table 94. Relative GPAM mRNA Levels in Primary Cyno Hepatocytes

Example 32: Screening additional human and cyno cross-reactive siRNAs in vitro in Hep3B cells, primary cyno hepatocytes, and primary human hepatocytes.

[0576] Therapeutic siRNAs were designed to target human GPAM and cynomolgus monkey GPAM sequence. The activities of the siRNAs were assessed in Hep3B cells using transfection reagent or in human hepatocytes using free uptake through the ASGPR. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 95, where Nf (e. g. , Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoromodified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 96.

[0577] Hep3B (ATCC, HB-8064) cells in EMEM + 10% FBS (Cytiva) were plated 10,000 cells per well in a 96- well plate and grown to -70% confluence. Transfection complexes were made by mixing 10 nM of siRNA in 25 μL of Opti-MEM media (Thermo Scientific) and 1.5 μL of Lipofectamine RNA-MAX (Fisher, 13778150) in 25 μL of Opti-MEM media together and incubating 5 minutes at room temperature. The siRNAs were prepared in triplicate and 10 μL of the transfection complexes was added to the cells. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828).

[0578] Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (Thermo Fisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 97. Of the siRNAs screened in Hep3Bs, most had some activity with transfection reagent and the best sequences were ETD03024, ETD03025, ETD03030, ETD03031, and ETD03033.

[0579] Primary human hepatocytes (BioIVT) were plated 50,000 cells per well in a 96 -well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty -four hours post thawing, hepatocytes were treated with lOul siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 98. Of the siRNAs screened in primary human hepatocytes, the best sequences were ETD03031, ETD03033-ETD03036.

[0580] Primary cyno hepatocytes (BioIVT) were plated 50,000 cells per well in a 96-well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty -four hours post thawing, hepatocytes were treated with 10 μL siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for cynomolgus monkey GPAM (ThermoFisher, assay# Mf02878271_ml) and cynomolgus monkey housekeeping gene GUSB (ThermoFisher, assay# Mf04392669_ml) and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222)). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 99. Of the siRNAs screened in primary cyno hepatocytes, the best sequences were ETD03024, ETD03025, and ETD0303 1.

Table 95. Example siRNA Sequences

Table 96. Example siRNA Base Sequences

Table 97. Relative GPAM mRNA Levels in Hep3B cells

Table 98. Relative GPAM mRNA Levels in Primary Human Hepatocytes

Table 99. Relative GPAM mRNA Levels in Primary Cyno Hepatocytes

Example 33: Screening additional human and cyno (1 mismatch) siRNAs in vitro in Hep3B cells, primary cyno hepatocytes, and primary human hepatocytes.

[0581] Therapeutic siRNAs were designed to target human GPAM and cynomolgus monkey with one mismatch with cynomolgus monkey GPAM sequence. The activities of the siRNAs were assessed in Hep3B cells using transfection reagent or in human or cyno primary hepatocytes using free uptake through the ASGPR. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 100, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluor o-modified nucleoside, n (e.g., a, c, g, t, or u) is a2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 101.

[0582] Hep3B (ATCC, HB-8064) cells in EMEM + 10% FBS (Cytiva) were plated 10,000 cells per well in a 96- well plate and grown to -70% confluence. Transfection complexes were made by mixing 10 nM of siRNA in 25 μL of Opti-MEM media (Thermo Scientific) and 1.5 μL of Lipof ectamine RNA-MAX (Fisher, 13778150) in 25 μL of Opti-MEM media together and incubating 5 minutes at room temperature. The siRNAs were prepared in triplicate and 10 μL of the transfection complexes was added to the cells. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828).

[0583] Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (Thermo Fisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 102. Of the siRNAs screened in Hep3Bs, most had some activity with transfection reagent and the best sequences were ETD03037, ETD03043-ETD03045, and ETD03049.

[0584] Primary human hepatocytes (BioIVT) were plated 50,000 cells per well in a 96 -well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty -four hours post thawing, hepatocytes were treated with lOul siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for human GPAM (ThermoFisher, assay# Hs01573684_ml) and the human housekeeping gene PPIA (ThermoFisher, assay# Hs99999904_ml), and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419- 222). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 103. Of the siRNAs screened in primary human hepatocytes, the best sequences were ETD03038, ETD03039, and ETD03044.

[0585] Primary cyno hepatocytes (BioIVT) were plated 50,000 cells per well in a 96-well plate in Invitrogro CP Medium (BioIVT). Media was changed 4 hours after plating and thawing. Twenty -four hours post thawing, hepatocytes were treated with 10 μL siRNA diluted in PBS and 90 μL Invitrogro CP media for 1 uM concentration of siRNA in triplicate for free uptake. RNA was collected from the cells 48 hours post transfection using a MagMax RNA Isolation kit (ABI, A27828). Preparation of cDNA was performed using qScript Ultra Supermix (VWR, 95217) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for cynomolgus monkey GPAM (ThermoFisher, assay# Mf02878271_ml) and cynomolgus monkey housekeeping gene GUSB (ThermoFisher, assay# Mf04392669_ml) and PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222)). Data were normalized to the mean GPAM mRNA level in the untreated cells alone wells. Results are shown in Table 104. Of the siRNAs screened in primary cyno hepatocytes, the best sequences were ETD03044, ETD03045, and ETD03046.

Table 100. Example siRNA Sequences

Table 101. Example siRNA Base Sequences

Table 102. Relative GPAM mRNA Levels in Hep3B cells

Table 103. Relative GPAM mRNA Levels in Primary Human Hepatocytes

Table 104. Relative GPAM mRNA Levels in Primary Cyno Hepatocytes

Example 34: Screening human and non-human primate cross- reactive siRNAs ETD03031, ETD03033, ETD03034, ETD03036, ETD03038, ETD03039, ETD03043 and ETD03044 in AAV8- TBG-GLuc-h-GPAM

[0586] The activities of siRNAs, namely ETD03031, ETD03033, ETD03034, ETD03036, ETD03038, ETD03039, ETD03043, ETD03044, were assessed in mice transiently expressing human GPAM with a secreted Gaussia luciferase (GLuc) tag. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Tables 95-96 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e. g. , a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e. g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Tables 100-101.

[0587] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μl of PBS for a total 30 μl injection volume by the retroorbital route on Day -14. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels. Mice with low levels of GLuc were excluded from the study (< 700 ng/mL).

[0588] On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice bled on Day 11 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 105

Table 105. Relative GLuc Protein Level

Example 35: Screening siRNAs with alternative modification patterns of ETD03036 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0589] The base sequences of ETD03036 were synthesized to generate siRNAs (ETD03222- ETD03230) with alternative modification patterns. The activities of the siRNAs were assessed in mice transiently expressing human GPAM with a secreted Gaussia luciferase (GLuc) tag. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Tables 95-96 and 107 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphor othioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Tables 100-101 and 106

[0590] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -14. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0591] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n =5). Mice were bled on Day 10 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 108.

Table 106. Example siRNA Base Sequences

Table 107. Example siRNA Sequences

Table 108. Relative GLuc Protein Level

Example 36. Screening siRNAs with alternative modification patterns of ETD03033 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0592] The base sequences of ETD03033 were synthesized to generate siRNAs (ETD03212- ETD03221) with alternative modification patterns. The activities of the siRNAs were assessed in mice transiently expressing human GPAM with a secreted Gaussia luciferase (GLuc) tag. The activities of siRNAs, namely ETD03033, ETD03212-ETD03221, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 110, where Nf (e. g. , Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e. g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 109.

[0593] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -15. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0594] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n =5). Mice were bled on Day 12 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 111.

Table 109. Example siRNA Base Sequences

Table 110. Example siRNA Sequences

Table 111. Relative GLuc Protein Level

Example 37. Screening siRNAs with alternative modification patterns of ETD03011 in mice transfected with AAV8-TBG-GLuc-h-GPAM [0595] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03011, ETD03231-ETD03235, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03011 are shown in Tables 90-91. The siRNA sequences that were used are shown in Table 113, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoromodified nucleoside, n (e.g., a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 112.

[0596] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL 1 of PBS for a total 30 μL injection volume by the retroorbital route on Day -16. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 4182-6372 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 3)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0597] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice bled on Day 12 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 114.

Table 112. Example siRNA Base Sequences

Table 114. Relative GLuc Protein Level

Example 38: Screening siRNAs with alternative modification patterns of ETD03031 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0598] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03031 and ETD03236-ETD03244 and ETD03257, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03031 are shown in Tables 95-96, respectively. The siRNA sequences that were used are shown in Table 116, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 115.

[0599] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -18. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0600] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 117.

Table 115. Example siRNA Base Sequences

Table 116. Example siRNA Sequences

Table 117. Relative GLuc Protein Level

Example 39: Screening siRNAs with alternative modification patterns of ETD02996 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0601] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD02996 and ETD03245-ETD03256 and ETD03258, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD02996 are shown in Tables 85-86, respectively. The siRNA sequences that were used are shown in Table 119, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 118.

[0602] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -19. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 4182-6372 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 3)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0603] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 120.

Table 118. Example siRNA Base Sequences

Table 119. Example siRNA Sequences

Table 120. Relative Glue Protein Level

Example 40: Screening siRNAs with alternative modification patterns of ETD03013 in mice transfected with AAV8-TBG-Gluc-h-GPAM

[0604] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03013 and ETD03126-ETD03136, ETD03147-ETD03149 and ETD03260-ETD03261 were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03013 is shown in Tables 90-91. The other siRNA sequences that were used are shown in Table 122 where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a2’ -fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2’ deoxynucleoside, and “s” is a phosphorothioate linkage. The base sequences for each siRNA, with and without the 3’ UU extension, are shown in Table 121.

[0605] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -13. The recombinant AAV8 contains the Gaussia luciferase (Glue) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-Gluc-h-GPAM (Fragment 2)). Circulating levels of the secreted Glue were measured in plasma by a Glue GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma Glue concentration. The mice were allocated into groups with similar average Glue levels.

[0606] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n = 4). Mice were bled on Day 11 after subcutaneous injection with whole blood processed to plasma. Glue levels in the plasma were measured with Glue GLOW assay and normalized relative to the PBS control group. Results are shown in Table 123.

Table 121. Example siRNA Base Sequences

Table 122. Example siRNA Sequences

Table 123. Relative GLuc Protein Level

Example 41: Screening human and non-human primate cross- reactive siRNAs ETD03024- ETD03036 in AAV8- TBG- GLuc-h- GPAM.

[0607] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03024-ETD03036 were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03013 and ETD03147-ETD03149 are shown in Tables 90 -91 and Tables 121-122, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro- modified nucleoside, n (e.g., a, c, g, t, or u) is a2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage.

[0608] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -19. ETD03024-ETD03029, ETD03031- ET3034, and ETD03036 used the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). ETD03030 and ETD03035 were used the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 2103- 4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0609] On Day 0, infected mice were given a subcutaneous injection of a single 40 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n = 4). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 124.

Table 124. Relative GLuc Protein Level

Example 42: Screening human and non-human primate cross- reactive siRNAs ETD03225, ETD03244, ETD03215, ETD03281, ETD03282, ETD03260, ETD03247, and ETD03234 in AAV8-TBG-GLuc-h-GPAM.

[0610] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03234, ETD03260, ETD03215, ETD03247, ETD03244 and ETD03225 were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03281 and ETD03282 are shown in Tables 125 and 126 The siRNA sequences for ETD03234, ETD03260, ETD03215, ETD03247, ETD03244 and ETD03225 are shown in Tables 112-113, Tables 121-122, Tables 109-110, Tables 115-116 (ETD03244 and ETD03247), and Tables 106-107 respectively, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a2’-fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’- O-methyl modified nucleoside, nm (e. g., am, cm, gm, tm, or urn) is a 2’ -O-(2-methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage.

[0611] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -17. ETD03225, ETD03244, ETD03215( ETD03281 and ETD03282) used the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). ETD03260 used the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). ETdO3247 and ETD03234 used the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 4182-6372 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 3)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0612] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg or 30 μg dose of a GalNAc-conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 127.

Table 125. Example siRNA Base Sequences

Table 126. Example siRNA Sequences

Table 127. Relative GLuc Protein Level

Example 43: Screening siRNAs with alternative modification patterns of ETD03013 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0613] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03013, ETD03260, ETD03293-ETD03297 were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03013, ETD03260, ETD03293-ETD03297 are shown in Tables 90-91, Tables 121-122 and Tables 128-129, respectively, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g., a, c, g, t, or u) is a 2’ -O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or urn) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage.

[0614] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -15. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0615] On Day 0, infected mice were given a subcutaneous injection of a single 40 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 130.

Table 128 Example siRNA Base Sequences

Table 129. Example siRNA Sequences

Table 130. Relative GLuc Protein Level

Example 44: Screening siRNAs with alternative modification patterns of ETD03013 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0616] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03013, ETD03260, ETD03130, and ETD03295 were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03013, ETD03260, ETD03295 are shown in Tables 90-91, Tables 121-122 and Tables 128-129 respectively, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoro-modified nucleoside, n (e.g, a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage.

[0617] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -11. These siRNAs were screened with the recombinant AAV8 containing the Gaussia luciferase (GLuc) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels. [0618] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg or 30 μg dose of a GalNAc-conjugated siRNA or PBS as vehicle control (n = 5). Mice were bled on Day 10, Day 18, Day 28, and Day 35 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 131.

Table 131. Relative GLuc Protein Level

Example 45: Screening human and non-human primate cross- reactive siRNAs ETD02983, ETD02991, ETD02996, ETD03011, ETD03013, ETD03035, and ETD03147-ETD03149 in AAV8- TBG-GLuc-h-GPAM.

[0619] The activities of siRNAs, namely ETD02983, ETD02991, ETD03013, ETD03035, and ETD03147-ETD03149 were assessed in mice transiently expressing human GPAM (2103-4282 bp) with a secreted Gaussia luciferase (Glue) tag. The activities of siRNAs, namely ETD02996 and ETD03011 were assessed in mice transiently expressing human GPAM (4182-6372 bp) with a secreted Gaussia luciferase (GLuc) tag.

[0620] The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD02983, ETD02991, ETD02996 are shown in Tables 85-86, sequences for ETD03011, ETD03013 are shown in Tables 90-91; sequences for ETD03035 is shown in Tables 95-96; sequences for ETD03038 is shown in Tables 100-101 and sequences for ETD03147-ETD03149 is shown in Tables 121-122; where “NT’ is a2’-fluoro-modified nucleoside, “n” is a 2’ -O-methyl modified nucleoside, nm (e.g. , am, cm, gm, tm, or urn) is a 2’-O-(2- methoxyethyl) modified nucleoside, and “s” is a phosphorothioate linkage.

[0621] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno-associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for atotal 30 μL injection volume by the retroorbital route on Day -14. The first recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). The second recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 4182-6372 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 3)).

[0622] Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels. Mice with low levels of GLuc were excluded from the study (< 400 ng/mL).

[0623] On Day 0, infected mice were given a subcutaneous injection of a single 100 μg dose of a GalN Ac- conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice were bled on Day 11 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 132.

Table 132. Relative GLuc Protein Level

Example: 46: Screening human and non-human primate cross- reactive siRNAs ETD03130 and

ETD03247 in cynomolgus monkeys

[0624] Three groups (n=4/group) of ≥3-year-old male cynomolgus monkeys were utilized for this study. The siRNA used in this Examples are included in Tables 121-122 (ETD03130), and Tables 118-119 (ETD03247), respectively where "Nf is a 2’-fluoro-modified nucleoside, "n" is a 2' O- methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, and "s" is a phosphorothioate linkage.

[0625] The 12 male cynomolgus monkeys were randomized and divided into 3 treatment groups, with body weights approximately balanced across groups, based on different test articles and dosage levels. On Day 0, Group 1 cynomolgus monkeys were injected subcutaneously (0.2mL/kg) with 10 mM PBS, Group 2 cynomolgus monkeys were injected subcutaneously (0.2mL/kg) with 2 mg/kg ETD03130 at a concentration of 10 mg/mL, and Group 3 cynomolgus monkeys were injected subcutaneously (0.2mL/kg) with 2 mg/kg ETD03247 at a concentration of 10 mg/mL.

[0626] Liver Biopsy:

[0627] On Day -8 and Day 28, animals were fasted overnight prior to the liver biopsy procedure. For the procedure, animals were appropriately anesthetized using a combination of ketamine (5-15 mg/kg, IM) and Dexmedetomidine (0.02 mg/kg, IM). Buprenorphine (0.24 mg/kg, SC) given once every 24 hours post-biopsy for 3 days for analgesia. For Direct Visualization, the incision site was shaved and disinfected with Povidone Iodine 10% solution (Betadine, Aspen Veterinary Resources, Liberty, MO, USA, for external animal use only.). A local anesthesia (Lidocaine HCl 2%, ~2 mg/kg) was administered in the incision area prior to liver biopsy procedure. A 1.5 cm abdominal incision was made using a sterile stainless steel disposable scalpel [10 (4-410) or 15 (4-415), Integra, Kai Medical], immediately caudal to the xiphoid process. The underlying musculature, if any, was split longitudinally to expose the peritoneum to be incised. The surgery incision was performed on the same midline incision or at the Principal Surgeon’s discretion. The incision will be held open with Gelpi’s retractor to visualize the liver. The 12g x 10 cm biopsy needle (Cat. # MQ1210, Bard Marquee Disposable Core Biopsy Instrument, Lot # 0001407900, Penetration Depth = 18 mm and 25 mm. Length of Sample Notch = 17 mm and 19 mm. Tempe, AZ 85281) was aligned in such a way as to collect as much of the liver as possible, while avoiding the hepatic veins, and activated to obtain the sample. The entry site of the needle was packed with a fiber-free gauze sponge, as well as the surrounding area to collect any blood loss because of the procedure. When hemostasis was achieved, the wound was closed with a cruciate-style suture pattern of 3-0 or 4-0 absorbable suture material [Coated Vicryl (Polyglactin 910), Ethicon], and the skin closed in a subcuticular pattern with similar material and glued together using atopical skin adhesive [2-Octyl Cyanoacrylate, Skinaffix, Medline], [0628] The biopsied liver tissue was then immediately placed in 500 μL cold RNAlater in a 2 mL cryovial. Cryovials were briefly spun to ensure tissue is fully submerged in RNAlater and stored for 24 hours at 4°C. After 24 hours, the RNAlater was removed from the tube then transferred into a freezer maintained at -80°C.

[0629] In addition to the post-surgical care described above, animals received Excede® (Zoetis) broad spectrum cephalosporin antibiotic (Ceftiofur Crystalline Free Acid, 6.6 mg/kg, SC) and Atipamezole (0.225 mg/kg IM). [0630] qRT-PCR: Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The relative levels of liver GPAM mRNA were assessed by RT-qPCR in triplicate on a QuantStudio™ 6 Pro Real- Time PCR System using TaqMan assays for cynomolgus monkey GPAM (ThermoFisher, assay# Mf02878271_ml), and the cynomolgus monkey housekeeping gene PPIB (ThermoFisher, assay# Mf02802985_ml) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data was normalized to the study Day -8 pre-dose liver biopsy for each individual by the delta-delta Ct method. The results of the Day 28 liver biopsy mRNA analysis are summarized in Table 133.

Table 133. Normalized Individual and Mean Level of Cynomolgus Monkey GPAM mRNA.

Example 47. EMPD-466 safety and tolerability study of ETD03215, ETD03130, ETD03247 and ETD03234 in C57B1/6 mice

[0631] The siRNAs were conjugated to the GalNAc ligand ETL17. The siRNAs, namely ETD03215, ETD03130, ETD03247 and ETD03234 were assessed for potential toxicity in mice. The siRNAs contained the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences for ETD03215, ETD03130, ETD03247 and ETD03234 are shown in Tables 109-110 (ETD03215), Tables 121-122 (ETD03130), Tables 118-119 (ETD03247), and Tables 112-113 (ETD03234), respectively, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a 2’ -fluoromodified nucleoside, n (e.g., a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2-methoxyethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, and “s” is a phosphorothioate linkage.

[0632] Six- to eight-week-old female (C57B1/6) mice were given a subcutaneous injection of 200 μg dose of a GalN Ac-conjugated siRNA or PBS as vehicle control (n ≥ 4) on Days 0, 4, and 8. Mice were bled on Days 2, 10, and 15 after subcutaneous injection with whole blood processed to serum for the following clinical chemistry assays: ALB, ALP, ALT, AST, TBIL, BUN, BUN:CREA Ratio, CK, CREA, GGT, GLOB, TP. Results are shown in Tables 134-136.

Example 48: Screening siRNAs with alternative modification patterns of ETD03247 in mice transfected with AAV8-TBG-GLuc-h-GPAM

[0633] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs, namely ETD03247 and ETD03553-ETD03556 and ETD03558- ETD03560, were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphorothioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 137, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a2’-fhroro-modified nucleoside, n (e.g, a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, [NUNA] is an unlocked nucleoside and “s” is a phosphorothioate linkage. The base sequences for each siRNA, without the 3’ UU extension, are shown in Table 138.

[0634] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno- associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for a total 30 μL injection volume by the retroorbital route on Day -22. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 4182-6372 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 3)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coel enter azine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0635] On Day 0, infected mice were given a subcutaneous injection of a single 100 μg or 40 μg dose of a GalNAc-conjugated siRNA or PBS as vehicle control (n ≥ 4). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 139.

Table 137: Modified Sequences

Table 138: Base Sequences

Table 139: Relative plasma levels of luciferase

Example 49: Therapeutic siRNA-mediated knockdown of GPAM in combination with a Glucagon- like peptide-1 receptor agonist (GLP-1RA) in a mouse model of non-alcoholic fatty liver disease (NAFLD) using a high fat, high fructose diet mouse model and a choline deficient high fat diet mouse model

[0636] High fat, high fructose diet (HFHFD)

[0637] The effects of siRNA-mediated knockdown of GPAM in the liver were investigated in a high fat, high fructose diet-based mouse model of non-alcoholic fatty liver disease (NAFLD). Six- to eight-week- old C57BL/6NHsd female mice (Envigo) were given normal chow (Groups 1 and 2) or a high fat diet (Envigo TD.120330, 0.2% cholesterol, 45% fat by calories) and high fructose water (55% fructose, 45% glucose) for 8 weeks (Groups 3-6) prior to siRNA injection.

[0638] After 8 weeks, mice in Group 1 on regular chow (n=20) and Group 3A on HFHFD (n=5) were injected with 100 μl of phosphate buffered saline (PBS) subcutaneously biweekly until 16 weeks (Day 112). Mice in Group 2 on regular chow (n=20) and Group 4 on HFHFD (n=10) were injected biweekly with a 3 mg/kg dose of ETD02284 (Table 140). [0639] After 8 weeks, mice in Group 3B (n=5) on HFHFD were injected daily with 100 μl of PBS subcutaneously and Group 5 on HFHFD (n=10) were injected with 100 μl of GLP1 -RA Semaglutide at a 0. 12 mg/kg dose subcutaneously daily until 16 weeks. This dose was escalated over 5 days at a rate 0.024, 0.048, 0.072, 0.096 mg/kg/day to the final dose of 0.12 mg/kg/day.

[0640] After 8 weeks, mice in Group 6 on HFHFD (n=10) were injected with 100 μl of Semaglutide at 0. 12 mg/kg dose subcutaneously daily and 100 μl of ETD02284 at 3 mg/kg dose subcutaneously biweekly until 16 weeks. The Semaglutide dose was escalated in this group in the same manner as Group 5.

Table 140. Dosing Groups for HFHFD

[0641] Body weights were recorded weekly at the start of the HFHFD until the end of the study. Mice were fasted 16 hours for serum collection biweekly from the start of study until 16 weeks (Day 112). Serum was assessed for the following clinical chemistry assays at IDEXX Laboratories, Incorporated - ALP, ALT, BUN, cholesterol, HDL, LDL, glucose, total bilirubin, total protein, triglycerides, and betahydroxybutyrate. Ketones were also measured in a drop of whole blood using Precision Xtra Ketone Test Strips (Abbott).

[0642] All mice in Groups 1-6 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 16 weeks. The liver weight was taken and then the liver was separated into 3 sections. One section was placed in RNAlater for qRT-PCR (Thermo Fisher #AM7020), one section was snap frozen in liquid nitrogen for liver triglycerides and liver collagen measurement, one section was placed into 10% formalin for fixation and paraffin embedding for H&E and Picrosirius Red staining for histopathology.

[0643] Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for mouse GPAM (ThermoFisher, assay# Mm01261106_ml), mouse COL1A1 (ThermoFisher, assay # MmOO8O I666_g1 ), mouse TIMP1 (ThermoFisher, assay # Mm00801666_gl), mouse TGFB1 (ThermoFisher, assay # Mm01341361_ml), mouse Acta2/SMA (ThermoFisher, assay # Mm00725412_sl), mouse Tnf-α (ThermoFisher, assay # Mm00443258_ml), mouse II IB (ThermoFisher, assay # Mm00434228_ml), mouse CCL2 (ThermoFisher, assay # Mm00441242_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm()2342430_gl ) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data was normalized to the mouse group receiving PBS (Normal Diet - Group 1, HFHFD - Group 3) by the delta-delta Ct method.

[0644] The 16 -week GPAM mRNA levels are shown in Table 141. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or HFHFD (Group 3). Mice on the normal diet treated with ETD02284 (Group 2) showed reduced liver GPAM liver mRNA levels compared to the PBS group (Group 1). Mice on the HFHFD treated with ETD02284 (Group 4 and 6) had reduced liver GPAM mRNA levels compared to mice on the HFHFD receiving PBS (Group 3).

Table 141. Liver GPAM mRNA Levels in HFHFD mice at 16 weeks.

[0645] The 16 -week pro-inflammatory genes CCL2 and TIMP1 mRNA levels are shown in Table 142. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or HFHFD (Group 3). Mice on the HFHFD treated with Semaglutide (Group 5) showed reduction of CCL2 and TIMP1 mRNA levels compared to mice on the HFHFD receiving PBS (Group 3). Mice on the HFHFD treated with Semaglutide and ETD02284 (Group 6) showed reduction of CCL2 and TIMP1 mRNA levels compared to mice on the HFHFD receiving PBS (Group 3).

Table 142. Liver CCL2 and TEMP1 mRNA Levels in HFHFD mice at 16 weeks.

[0646] The 16 -week pro-fibrotic genes TGFβ1, ACTA2/SMA, COL1A1, and COL3A1 mRNA levels are shown in Table 143. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or HFHFD (Group 3). Mice on the normal diet treated with ETD02284 (Group 2) showed a reduction of TGFβ1, ACTA2, and COL3A1 liver mRNA levels compared to the PBS group (Group 1). Mice on the HFHFD treated with ETD02284 (Group 4) had reduced TGFpi liver mRNA levels compared to mice on the HFHFD receiving PBS (Group 3). Mice on the HFHFD treated with Semaglutide (SM) (Group 5) showed reduced ACTA2 mRNA levels compared to mice on the HFHFD receiving PBS (Group 3). Mice on the HFHFD treated with Semaglutide (SM) and ETD02284 (Group 6) showed reduced COL1 Al and COL3A1 mRNA levels compared to mice on the HFHFD receiving PBS (Group 3).

Table 143. Liver TGFpi, ACTA2/SMA, COL1A1, and COL3A1 mRNA Levels in HFHFD mice at

16 weeks

[0647] The ending body weights, body weight gain, and liver weight/body weight (LW/BW) ratio are shown in Table 144 at 16 weeks. Mice on the HFHFD (Groups 3-6) showed an increase in weight gained and increase in the LW/BW ratio compared to mice on normal diet. Mice on the HFHFD treated with Semaglutide (Group 5) showed a lower body weight at 16 weeks compared to the PBS group (Group 3). Mice on the HFHFD treated with Semaglutide and ETD02284 (Group 6) showed lowered body weight and reduced body weight gain at 16 weeks compared to the PBS group (Group 3).

Table 144. Body weight, body weight gained and LW/BW ratios of HFHFD mice at 16 weeks

[0648] The 16-week serum levels of ALT, ALP, and TBIL are shown in Table 145. Mice on the HFHFD treated with Semaglutide (Group 5) showed reduction of ALT at 16 weeks compared to the PBS group (Group 3). Mice on the HFHFD treated with Semaglutide and ETD02284 (Group 6) showed reduction of ALT at 16 weeks compared to the PBS group (Group 3). Table 145. ALP, ALT, and TBIL serum levels of HFHFD mice at 16 weeks.

[0649] The 16-week serum levels of CHOL, HDL, and LDL are shown in Table 146. In mice on normal diet, treatment with ETD02284 (Group 2) had reduced CHOL and HDL levels compared to the PBS group (Group 1). Mice on the HFHFD treated with Semaglutide and ETD02284 (Group 6) showed reduction of CHOL and HDL at 16 weeks compared to the PBS group (Group 3).

Table 146. CHOL, HDL, and LDL serum levels of HFHFD mice at 16 weeks.

[0650] The 16-week serum levels of BHB are shown in Table 147. In mice on normal diet, treatment with ETD02284 (Group 2) had increased BHB serum levels compared to the PBS group (Group 1). Mice on the HFHFD treated with Semaglutide (Group 5) showed a reduction of BHB serum levels at 16 weeks compared to the PBS group (Group 3).

Table 147. BHB serum levels of HFHFD mice at 16 weeks.

[0651] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride Colorimetric Kit (Cayman Chemical #10010303) following the manufacturer’s instructions. The relative levels of triglycerides in the flash frozen liver sections are shown in Table 148 with the data normalized to mice on normal diet treated with PBS (Group 1) or mice on the HFHFD treated with PBS (Group 3). In mice on normal diet, treatment with ETD02284 (Group 2) had decreased liver triglyceride levels compared to the PBS group (Group 1). Mice on the HFHFD treated with ETD02284 (Group 4) showed reduction of liver triglyceride levels at 16 weeks compared to the PBS group (Group 3). Mice on the HFHFD treated with Semaglutide and ETD02284 (Group 6) showed reduction of liver triglyceride levels at 16 weeks compared to the PBS group (Group 3).

Table 148. Triglyceride levels in the liver of HFHFD mice at 16 weeks.

[0652] Liver hydroxyproline levels were measured in a piece of flash frozen liver as surrogate for liver collagen levels using a Hydroxyproline Assay Kit (Abeam ab222941) following the manufacturer’s instructions. The levels of liver hydroxyproline in the flash frozen liver sections are shown in Table 149. Mice on the HFHFD treated with the combination of Semaglutide and ETD02284 (Group 6) showed reduction of liver hydroxyproline compared to the PBS group (Group 3).

Table 149. Hydroxyproline/Collagen levels in the liver of HFHFD mice at 16 weeks.

[0653] To assess liver histology, the formalin fixed livers were paraffin embedded, sectioned, and then stained with hematoxylin and eosin (H&E). Steatotic quantification of the H&E-stained livers utilized an algorithm to quantify lipid droplet area within the tissue section and reported as % lipid area, % steatotic area, and % microsteatotic area of the liver section for each animal. These results are summarized in Table 150. In mice on normal diet, treatment with ETD02284 (Group 2) had decreased liver % lipid area and % steatotic area compared to the PBS group (Group 1). Mice on the HFHFD treated with ETD02284 (Group 4), Semaglutide (Group 5), and the combination of Semaglutide and ETD02284 (Group 6) showed reduction of liver % lipid area, % steatotic area, and % microsteatotic area compared to the PBS group (Group 3). Table 150. Percent (%) Steatotic Area of HFHFD Livers at 16 weeks.

[0654] Choline-Deficient, L-amino Acid-Defined, High-Fat Diet (CDAHFD)

[0655] The effects of siRNA-mediated knockdown of GPAM in the liver was investigated in a Choline- Deficient, L-amino Acid-Defined, High-Fat Diet-based mouse model of non-alcoholic fatty liver disease (NAFLD). Six- to eight-week-old C57BL/6NHsd female mice (Envigo) were given normal chow (Groups 1 and 2) or ahigh fat diet (Envigo TD. 120330, 0.2% cholesterol, 45% fat by calories) and high fructose water (55% fructose, 45% glucose) for 4 weeks (Groups 3-6). Groups 3-6 were then given a choline deficient high fat diet (Research Diets A06071302, L-amino Acid Diet With 60 kcal% Fat With 0. 1% Methionine and No Added Choline) for 12 weeks.

[0656] After 8 weeks, mice in Group 1 on regular chow (n=20) and Group 3A on CDAHFD (n=5) were injected with 100 μl of phosphate buffered saline (PBS) subcutaneously biweekly until 16 weeks (Day 112). Mice in Group 2 on regular chow (n=20) and Group 4 on CDAHFD (n=10) were injected biweekly with a 3 mg/kg dose of ETD02284 (Table 151).

[0657] After 8 weeks, mice in Group 3B (n=5) on CDAHFD were injected daily with 100 pl of PBS subcutaneously and Group 5 on CDAHFD (n=10) were injected with 100 μl of GLP-1RA Semaglutide at a 0. 12 mg/kg dose subcutaneously daily until 16 weeks. This dose was escalated over 5 days at a rate 0.024, 0.048, 0.072, 0.096 mg/kg/day to the final dose of 0. 12 mg/kg/day.

[0658] After 8 weeks, mice in Group 6 on CDAHFD (n=10) were injected with 100 μl of Semaglutide at 0. 12 mg/kg dose subcutaneously daily and 100 μl of ETD02284 at 3 mg/kg dose subcutaneously biweekly until 16 weeks. The Semaglutide dose was escalated in this group in the same manner as Group 5.

Table 151. Dosing Groups for CDAHFD

[0659] Body weights were recorded weekly until the end of the study. Mice were fasted 16 hours for serum collection biweekly from the start of study until 16 weeks (Day 112). Serum was assessed for the following clinical chemistry assays at IDEXX Laboratories, Incorporated - ALP, ALT, BUN, cholesterol, HDL, LDL, glucose, total bilirubin, total protein, triglycerides, and beta-hydroxybutyrate. Ketones were also measured in a drop of whole blood using Precision Xtra Ketone Test Strips (Abbott).

[0660] All mice in Groups 1-6 were euthanized via cervical dislocation following isoflurane exposure with a final serum bleed at 16 weeks. The liver weight was taken and then the liver was separated into 3 sections. One section was placed in RNAlater for qRT-PCR (Thermo Fisher #AM7020), one section was snap frozen in liquid nitrogen for liver triglycerides and liver collagen measurement, one section was placed into 10% formalin for fixation and paraffin embedding for H&E and Picrosirius Red staining for histopathology.

[0661] Total liver RNA was prepared by homogenizing the RNAlater liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations. Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions. The levels of liver GPAM mRNA were assessed by RT- qPCR in triplicate on a QuantStudio™ 6 Pro Real-Time PCR System using TaqMan assays for mouse GPAM (ThermoFisher, assay# Mm01261106_ml), mouse COL1A1 (ThermoFisher, assay # Mm00801666_g I ). mouse TIMP1 (ThermoFisher, assay # Mm00801666_gl), mouse TGFB1 (ThermoFisher, assay # Mm01341361_ml), mouse Acta2/SMA (ThermoFisher, assay # Mm00725412_sl), mouse Tnf-α (ThermoFisher, assay # Mm00443258_ml), mouse II 1 B (ThermoFisher, assay # Mm00434228_ml), mouse CCL2 (ThermoFisher, assay # Mm00441242_ml), and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm( >234243 ()_g I ) using PerfeCTa® qPCR FastMix®, Low ROX™ (VWR, Catalog# 101419-222). Data was normalized to the mouse group receiving PBS (Normal Diet - Group 1, CDAHFD - Group 3) by the delta-delta Ct method.

[0662] The 16 -week GPAM mRNA levels are shown in Table 152. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or CDAHFD (Group 3). Mice on the normal diet treated with ETD02284 (Group 2) showed reduced liver GPAM liver mRNA levels compared to the PBS group (Group 1). Mice on the CDAHFD treated with ETD02284 (Group 4 and 6) had reduced liver GPAM mRNA levels compared to mice on the CDAHFD receiving PBS (Group 3).

Table 152. Liver GPAM mRNA Levels in CDAHFD mice at 16 weeks.

[0663] The 16 -week pro-inflammatory genes CCL2 and TIMP1 mRNA levels are shown in Table 153. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or CDAHFD (Group 3). Mice on the CDAHFD treated with ETD02284 (Group 4) showed reduction of CCL2 and TIMP1 mRNA levels compared to mice on the CDAHFD receiving PBS (Group 3). Mice on the CDAHFD treated with Semaglutide (Group 5) showed a reduction of CCL2 and TIMP1 mRNA levels compared to mice on the CDAHFD receiving PBS (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed a reduction of CCL2 and TIMP1 mRNA levels compared to mice on the CDAHFD receiving PBS (Group 3).

Table 153. Liver CCL2 and TIMP1 mRNA Levels in CDAHFD mice at 16 weeks.

[0664] The 16 -week pro-fibrotic genes TGFpi, ACTA2/SMA, COL1A1, and COL3A1 mRNA levels are shown in Table 154. Data were normalized to the level in animals receiving PBS on normal diet (Group 1) or CDAHFD (Group 3). Mice on the normal diet treated with ETD02284 (Group 2) showed a reduction of TGFpi, ACTA2, and COL3A1 liver mRNA levels compared to the PBS group (Group 1). Mice on the CDAHFD treated with ETD02284 (Group 4) showed a reduction of ACTA2/SMA, COL1 Al and COL3A1 compared to mice on the CDAHFD receiving PBS (Group 3). Mice on the CDAHFD treated with Semaglutide (Group 5) showed a reduction of TGFpi mRNA compared to mice on the CDAHFD receiving PBS (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed a reduction to TGFpi, COL1A1, and COL3A1 and ACTA2/SMA compared to mice on the CDAHFD receiving PBS (Group 3).

Table 154. Liver TGFpi, ACTA2/SMA, COL1A1, and COL3A1 mRNA Levels in CDAHFD mice at

16 weeks.

[0665] The ending body weights, body weight gain, and liver weight/body weight (LW/BW) ratio are shown in Table 155 at 16 weeks. Mice on the CDAHFD (Groups 3-6) showed an increase in weight gained and an increase in the LW/BW ratio compared to mice on normal diet. Mice on the CDAHFD treated with ETD02284 (Group 4) showed a reduction of the LW/BW ratio at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide (Group 5) showed a lower body weight and reduced weight gain and an increase in the LW/BW ratio at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed lowered body weight, reduced body weight gain, and a reduced LW/BW ratio at 16 weeks compared to the PBS group (Group 3).

Table 155. Body weight, body weight gained and LW/BW ratios of CDAHFD mice at 16 weeks.

[0666] The 16-week serum levels of ALT, ALP, and TBIL are shown in Table 156. Mice on the CDAHFD (Groups 3-6) showed increases in ALP, ALT, and TBIL compared to mice on normal diet. Mice on the CDAHFD treated with ETD02284 (Group 4) showed a reduction of ALP, ALT, and TBIL at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide (Group 5) showed a reduction of ALT at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed a reduction of ALT at 16 weeks compared to the PBS group (Group 3).

Table 156. ALP, ALT, and TBIL serum levels of CDAHFD mice at 16 weeks.

[0667] The 16-week serum levels of CHOL, HDL, and LDL are shown in Table 157. In mice on normal diet, treatment with ETD02284 (Group 2) had reduced CHOL and HDL levels compared to the PBS group (Group 1). Mice on the CDAHFD treated with Semaglutide (Group 5) showed reduction CHOL, HDL, and LDL at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed reduction of CHOL, HDL, and LDL at 16 weeks compared to the PBS group (Group 3).

Table 157. CHOL, HDL, and LDL serum levels of CDAHFD mice at 16 weeks.

[0668] The 16-week serum levels of BHB are shown in Table 158. In mice on normal diet, treatment with ETD02284 (Group 2) increased BHB serum levels compared to the PBS group (Group 1). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed an increase BHB serum levels at 16 weeks compared to the PBS group (Group 3).

Table 158. BHB serum levels of CDAHFD mice at 16 weeks.

[0669] Liver triglycerides were measured in a piece of flash frozen liver using a Triglyceride Colorimetric Kit (Cayman Chemical #10010303) following the manufacturer’s instructions. The relative levels of triglycerides in the flash frozen liver sections are shown in Table 159 with the data normalized to mice on normal diet treated with PBS (Group 1) or mice on the CDAHFD treated with PBS (Group 3). In mice on normal diet, treatment with ETD02284 (Group 2) decreased liver triglyceride levels compared to the PBS group (Group 1). Mice on the CDAHFD treated with Semaglutide (Group 5) showed a reduction of liver triglyceride levels at 16 weeks compared to the PBS group (Group 3). Mice on the CDAHFD treated with Semaglutide and ETD02284 (Group 6) showed a reduction of liver triglyceride levels at 16 weeks compared to the PBS group (Group 3).

Table 159. Relative Triglyceride levels in the liver of CDAHFD mice at 16 weeks.

[0670] Liver hydroxyproline levels were measured in a piece of flash frozen liver as surrogate for liver collagen levels using a Hydroxy proline Assay Kit (Abeam ab222941) following the manufacturer’s instructions. The liver levels of hydroxyproline in the flash frozen liver sections are shown in Table 160. Mice on the CDAHFD treated with ETD02284 (Group 4) and the combination of ETD02284 and Semaglutide (Group 6) showed reduced liver hydroxyproline levels at 16 weeks compared to the PBS group (Group 3).

Table 160. Hydroxyproline/Collagen levels in the liver of CDAHFD mice at 16 weeks.

[0671] To assess liver histology, the formalin fixed livers were paraffin embedded, sectioned, and then stained with hematoxylin and eosin (H&E). Steatotic quantification of the H&E-stained livers utilized an algorithm to quantify lipid droplet area within the tissue section and reported as % lipid area, % microsteatotic area, and % steatotic area of the liver section for each animal. These results are summarized in Table 161. Mice on the CDAHFD treated with ETD02284 (Group 4) and the combination of ETD02284 and Semaglutide (Group 6) showed a reduction in % lipid area and % steatotic area compared to the PBS group (Group 3).

Table 161. Percent (%) Steatotic Area of CDAHFD Livers at 16 weeks.

[0672] Picrosirius red staining for collagen in the CDAHFD showed increased levels of parenchymal collagen. This picrosirius red staining in the parenchyma was quantified using an algorithm that filtered out blood vessels and structural collagen present in the liver section and then reported as % area of the liver section and the results are shown in Table 162. Mice on the CDAHFD treated with ETD02284 (Group 4), Semaglutide (Group 5), and the combination of Semaglutide and ETD02284 (Group 6) showed a reduction in % picrosirius red staining in the parenchyma compared to the PBS group (Group 3).

Table 162. % PSR Parenchymal Area of CDAHFD Livers at 16 weeks.

Example 50: Screening human and non-human primate-cross-reactive siRNAs ETD03613- ETD03624 in AAV8-TBG-GLuc-h-GPAM

[0673] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 164, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a2’-fluoro-modified nucleoside, n (e.g, a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, [NUNA] is an unlocked nucleoside and “s” is a phosphorothioate linkage. The base sequences for each siRNA, without the 3’ UU extension, are shown in Table 163.

[0674] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno- associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for a total 30 μL injection volume by the retroorbital route on Day -21. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 1-2180 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 1)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1 : 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coelenterazine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0675] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n =5). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 165. Table 163. Example siRNA Base Sequences

Table 164. Example siRNA Sequences

Table 165. Relative Plasma Levels of Gaussia Luciferase

Example 51: Screening human and non-human primate-cross-reactive siRNAs ETD03625- ETD03627 in AAV8-TBG-GLuc-h-GPAM.

[0676] The siRNAs are conjugated to the GalNAc ligand ETL17. The activities of siRNAs were assessed in mice transiently expressing human GPAM. The siRNAs contain the GalNAc ligand ETL17 followed by a phosphor othioate linkage at the 5’ end of the sense strand. The siRNA sequences that were used are shown in Table 167, where Nf (e.g., Af, Cf, Gf, Tf, or Uf) is a2’-fluoro-modified nucleoside, n (e.g, a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, nm (e.g., am, cm, gm, tm, or um) is a 2’-O-(2- methoxy ethyl) modified nucleoside, dN (e.g., dA, dC, dG, dT, or dU) is a 2' deoxynucleoside, [NUNA] is an unlocked nucleoside and “s” is a phosphorothioate linkage. The base sequences for each siRNA, without the 3’ UU extension, are shown in Table 166.

[0677] Six- to eight-week-old female mice (C57B1/6) were injected with 10 μL of a recombinant adeno- associated virus 8 (AAV8) vector (≥1.2 x 10¹³ genome copies/mL) and 20 μL of PBS for a total 30 μL injection volume by the retroorbital route on Day -21. The recombinant AAV8 contains the Gaussia luciferase (GLuc) tag and positions 2103-4282 of the open reading frame of the human GPAM sequence (ENST00000348367) under the control of the human thyroxine binding globulin (TBG) promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-GLuc-h-GPAM (Fragment 2)). Circulating levels of the secreted GLuc were measured in plasma by a GLuc GLOW assay (Nanolight Technology, #320-50). Briefly, recombinant Gaussia luciferase protein was diluted in a series to create a standard curve. Whole blood was collected from mice injected with AAV8, processed to plasma, and diluted 1: 10 in the GLOW assay buffer. The standard curve and samples were then incubated with coel enter azine substrate for 5 minutes then luminescence was measured on a plate reader (Perkin Elmer). Samples were interpolated based on the standard curve values for plasma GLuc concentration. The mice were allocated into groups with similar average GLuc levels.

[0678] On Day 0, infected mice were given a subcutaneous injection of a single 60 μg dose of a GalNAc- conjugated siRNA or PBS as vehicle control (n =5). Mice were bled on Day 14 after subcutaneous injection with whole blood processed to plasma. GLuc levels in the plasma were measured with GLuc GLOW assay and normalized relative to the PBS control group. Results are shown in Table 168.

Table 166. Example siRNA Sequences

Table 167. Example siRNA Base Sequences

Table 168. Relative Plasma Levels of Gaussia Luciferase

[0679] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and compositions within the scope of these claims and their equivalents be covered thereby.

IV. SEQUENCE INFORMATION

[0680] Some embodiments include one or more nucleic acid sequences in the following table:

Table 169. Sequence Information

Table 170. Additional Sequence

Claims

CLAIMS What is claimed is:

1. A composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount decreases circulating cholesterol, apolipoprotein B, bilirubin, alanine aminotransferase, aspartate aminotransferase, or alkaline phosphatase in a subject, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69AS.

2. The composition of claim 1, wherein the cholesterol comprises total cholesterol, low density lipoprotein cholesterol, or non-high density lipoprotein cholesterol.

3. The composition of claim 1, wherein the cholesterol is decreased by about 10% or more, as compared to prior to administration.

4. A composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount decreases a liver fibrosis score, non-alcoholic fatty liver disease (NAFLD) activity score, or liver fat percentage in a subject, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69AS.

5. The composition of claim 4, wherein the decrease is by about 10% or more, as compared to prior to administration.

6. A composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount decreases a use of statin (HMG CoA reductase inhibitor) medication, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69AS.

7. The composition of claim 6, wherein the decrease is by about 10% or more, as compared to prior to administration.

8. A composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount improves a measurement that reflects a phenotype of esophageal varices, portal hypertension, NAFLD, NASH, alcoholic liver disease, liver fibrosis, liver cirrhosis, hepatocellular carcinoma, hyperlipidemia, ischemic heart disease, or coronary heart disease in a subject, wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 60S, 61S, 55S, 56AS, 9AS, 57AS, 30AS, 58AS, 68AS, and 69AS.

9. The composition of claim 8, wherein the improvement is by about 10% or more, as compared to prior to administration.

10. A composition comprising an siRNA that targets GPAM and when administered to a subject in an effective amount increases circulating ketone bodies in a subject wherein the siRNA comprises a modification pattern selected from the group consisting of 54S, 53S, 57S, 56S, 109S, 114S, 52S, 157S, 158S, 159S, 160S, 161S, 162S, 163S, 164S, 68AS, 69AS, 70AS, 71AS, and 72AS.

11. The composition of claim 10, wherein the increase is by about 10% or more, as compared to prior to administration.

12. The composition of any one of claims 1-11, wherein the oligonucleotide comprises an N- acetylgalactosamine (GalNAc) moiety, anN-acetylglucosamine (GlcNAc) moiety, or a mannose moiety, attached at a 3’ or 5’ terminus of the oligonucleotide.

13. The composition of any one of claims 1-12, wherein the oligonucleotide comprises a GalNAc moiety.

14. The composition of claim 13, comprising:

, wherein J comprises the oligonucleotide, and wherein J comprises an optional phosphate or phosphorothioate linking to the oligonucleotide.

15. The composition of any one of claims 1-14, wherein the siRNA comprises a sense strand and an antisense strand.

16. The composition of claim 15, wherein the sense strand comprises a sequence comprising at least 19 nucleosides of any one of SEQ ID NO: 1-6354, 13082-13402, 13951-14078, or 14285-14296, or 14337-14339.

17. The method of claim 15 or 16 wherein the antisense strand comprises a sequence comprising at least 19 nucleosides of any one of SEQ ID NO: 6355-12708, 13403-13723, 14079-14206, 14297- 14307, or 14340-14342.

18. A composition comprising an oligonucleotide that inhibits the expression of GP AM, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 12-30 nucleosides in length, wherein the siRNA comprises sense sequence comprising any one of SEQ ID NO: 13780-13840, 13914-13937, 14254-14255, 14308-14319, or 14331- 14333or an antisense sequence comprising any one of SEQ ID NO: 13841-13913, 13938-13950, 14207- 14253, 14256-14276, 14277-14284, 14320-14330, or 14334-14336.

19. The composition of any one of claims 1-18, further comprising a pharmaceutically acceptable carrier.

20. A method of treating a subject having liver disease, comprising administering an effective amount of the composition of claim 19 to the subject.

21. The method of claim 20, wherein the liver disease comprises NAFLD, NASH, alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

22. A method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition of claim 19 to the subject.

23. The method of claim 22, wherein the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease.

24. A method of treating a subject having liver disease, the method comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist.

25. The method of claim 24, wherein the liver disease comprises NAFLD (or MASLD), NASH (or MASH), alcoholic liver disease, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

26. The method of claim 24, wherein the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, dulaglutide, tirzepatide, dulaglutide, semaglutide or a combination thereof.

27. A method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist.

28. The method of claim 27, wherein the cardiometabolic disease comprises hyperlipidemia, ischemic heart disease, or coronary heart disease.

29. The method of claim 27, wherein the GLP-1 receptor agonist comprises exenatide, lixisenatide, liraglutide, dulaglutide, tirzepatide, dulaglutide, semaglutide or a combination thereof.

30. A composition comprising an oligonucleotide that inhibits the expression of GP AM in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 12-30 nucleosides in length, wherein the siRNA comprises sense sequence comprising any one of SEQ ID NO: 13780-13840, 13914-13937, 14254-14255, 14308-14319, or 14331- 14333or an antisense sequence comprising any one of SEQ ID NO: 13841-13913, 13938-13950, 14207- 14253, 14256-14276, 14277-14284, 14320-14330, or 14334-14336.

31. A method of treating a subject having liver disease, the method comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, glucose-dependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor-15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator -activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activin E expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof.

32. A method of treating a subject having cardiometabolic disease, comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one GLP-1 receptor agonist, glucose-dependent insulinotropic polypeptide (GIP) receptor agonist, GIP receptor antagonist, glucagon receptor agonist, amylin receptor agonist, apelin receptor agonist, peptide YY receptor agonist, calcitonin receptor agonist, growth differentiation factor-15 (GDF15) analogue, fibroblast growth factor 21 (FGF21) analog, fibroblast growth factor 21 (FGF19) analog, peroxisome proliferator -activated receptors (PPAR) agonist, thyroid hormone receptor-β agonist, FXR agonist, antagonist or modulator of inhibin βE (INHBE)/activin E expression or function, antagonist or modulator of activin receptor-like kinase 7 (ALK7) expression or function, antagonist or modulator of diacylglycerol acyltransferase (DGAT) or acetyl-CoA carboxylase (ACC) expression or function, antagonist or modulator of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression or function, antagonist or modulator of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression or function, antagonist or modulator of mitochondrial amidoxime-reducing component 1 (MTARC1) expression or function, or a combination thereof.

33. A method of treating a subject having liver or cardiometabolic disease, comprising administering an effective amount of the composition of claim 19 to the subject in combination with therapeutically effective amount of at least one additional active agent.