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WO2016072969A1 - Procédés de détection d'une chaîne lourde d'immunoglobuline et leurs procédés d'utilisation - Google Patents

Procédés de détection d'une chaîne lourde d'immunoglobuline et leurs procédés d'utilisation Download PDF

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WO2016072969A1
WO2016072969A1 PCT/US2014/063719 US2014063719W WO2016072969A1 WO 2016072969 A1 WO2016072969 A1 WO 2016072969A1 US 2014063719 W US2014063719 W US 2014063719W WO 2016072969 A1 WO2016072969 A1 WO 2016072969A1
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multiple myeloma
abundance
dominant
subject
active multiple
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Erming Tian
Joshua Epstein
Caleb STEIN
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6841—In situ hybridisation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/56—Staging of a disease; Further complications associated with the disease

Definitions

  • the present invention relates to a predictive model to determine treatment for subjects diagnosed with asymptomatic multiple myeloma.
  • MM Multiple myeloma
  • MM is a heterogeneous plasma cell disorder characterized by genetic abnormalities, including chromosomal translocations, deletions, duplications and genetic mutations.
  • MGUS Monoclonal gammopathy of undetermined significance
  • SMM multiple myeloma
  • the present disclosure encompasses a method of developing a predictive model to determine the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in-situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from each healthy individual is plotted as a point on an X/Y plot generating a normal clonal
  • the present disclosure encompasses a method to determine the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in- situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from each healthy individual is plotted as a point on an X/Y plot generating a normal clonal distribution; (d) calculating
  • the present disclosure encompasses a method of determining the treatment of a subject with pre-active multiple myeloma.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in-situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from each healthy individual is plotted as a point on an X/Y plot generating a normal clonal distribution; (d) calculating a region that encompasses the normal clon
  • abundance of the dominant Lambda chain clone from the subject diagnosed with pre- active multiple myeloma is plotted as a point on an X/Y plot; and (h) determining if the plotted point from the subject diagnosed with pre-active multiple myeloma is inside or outside the region, wherein (i) if the point is inside the region, the subject is at low risk of progressing to symptomatic multiple myeloma and is not treated and (ii) if the point is outside the region, the subject is at high risk of progressing to symptomatic multiple myeloma and is treated.
  • the present disclosure encompasses a method of determining the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining a biological sample from a subject diagnosed with pre-active multiple myeloma; (b) performing fluorescence in-situ hybridization (FISH) on the biological sample from the subject diagnosed with pre-active multiple myeloma using the same probes as used to develop the predictive model of the invention; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone from the subject diagnosed with pre-active multiple myeloma, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from the subject diagnosed with pre-active multiple myeloma is plotted as a point on the predictive model of the invention; and (d) determining the location
  • FIG. 1 depicts graphs showing a 100(1 -a)% prediction ellipse was based on normal distribution of clone diversities among healthy individuals (A) to stratify AMG (asymptomatic monoclonal gammopathy) and PPCD (polyclonal plasma cell dyscrasias) subgroups from MGUS (B), AMM (SMM) (C), and MM (D) diagnosed individuals.
  • AMG asymptomatic monoclonal gammopathy
  • PPCD polyclonal plasma cell dyscrasias
  • the methodology uses fluorescence in-situ hybridization (FISH) to identify low- and high-risk pre-active MM patients.
  • FISH fluorescence in-situ hybridization
  • the predictive model also allows follow-up analysis to determine changes in risk and response to therapy. It may also be used to identify early relapses in MM patients that achieved a complete response (CR) following therapy and thus the need for immediate intervention.
  • the methodology requires a minimal amount of biological sample without the need for purification of plasma cells from the biological sample.
  • the predictive model described herein is used to determine the treatment of a subject with pre-active multiple myeloma. I. FISH probe
  • a probe provided by the invention encompasses, in certain embodiments, a nucleic acid suitable for use as a fluorescent in situ hybridization (FISH) probe.
  • FISH is a cytogenetic technique used to visualize labeled DNA probes hybridized to a region of interest on a chromosome.
  • FISH probes are designed to only bind those parts of the chromosome with which they show a high degree of sequence complementarity. Accordingly, as used to refer to probes herein, the term
  • complementary includes “substantially complementary”, and refers to the ability of nucleic acids to hybridize by Watson-Crick base-pairing and form, at least partially, a double stranded structure.
  • a probe that hybridizes to a region of interest on a chromosome e.g., by Watson-Crick base-pairing
  • Watson-Crick base-pairing e.g., by Watson-Crick base-pairing
  • complementary means two sequences (e.g., a probe and a target sequence) hybridize under highly stringent hybridization conditions. "Highly stringent hybridization"
  • conditions in some embodiments, refers to under pH scale 7.0 at least about 50% (v/v) Formamide with 2X SSC and 0.1 % detergent at 42 °C, wth a first wash for 30 minutes at about 50 °C with about 50% (v/v) Formamide in 2X SSQ and with a subsequent wash with 1 XPBS and 0.01 % IGEPAL CA-630 at 37°C.
  • highly stringent hybridization conditions refers to at least about 6X SSC and 1 % SDS at 65 °C, with a first wash for 10 minutes at about 42 °C with about 20% (v/v) Formamide in 0.1 X SSC, and with a subsequent wash with 0.2 X SSC and 0.1 % SDS at 65 °C.
  • complementarity or substantially complementary means a nucleic acid sequence is at least about: 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99%, or more (e.g. 100%) identical to the complement (i.e. reverse complement) of a region of interest.
  • Reference to a pair of probes such as a pair of FISH probes, encompasses not only exactly two probes, such as a first probe and a second probe, but also, unless clearly indicated otherwise, additional probes, such as an additional 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, or more probes.
  • a probe of the invention provides one or more labeled DNA probes.
  • Labeled nucleic acid probes are synthetically and/or artificially labeled and are not naturally-occurring, but rather are products of human innovation.
  • Probes may be indirectly labeled with haptens or directly labeled with a fluorochrome- conjugated nucleotide, i.e., dUTP (2 -Deoxyuridine, 5 '-Triphosphate) in place of dTTP.
  • suitable haptens include biotin and digoxigenin.
  • Non-limiting examples of suitable fluorochromes include the Alexa fluor dye series, amino-methyl coumarin, cascade blue, cyanine 2 d , cyanine 3 d , cyanine 5 d , cyanine 7 d , DEAC, fluorescein, fluorescein isothiocyanate, rhodamine B and rhodamine derivatives (for example, 5(6)-carboxyrhodamine, 5-ROX, 5- TAMRA (5-carboxytetramethylrhodime), 6- TAMRA, etc.) and Texas Red.
  • the probe is directly labeled with a fluorochrome-conjugated nucleotide selected from the group consisting of DEAC- dUTP, 5-Fluorecein dUTP, 5(6)-Carboxyrhodamine 6G-dUTP, 5-TAMRA-dUTP, and 5- ROX-dUTP.
  • a fluorochrome-conjugated nucleotide selected from the group consisting of DEAC- dUTP, 5-Fluorecein dUTP, 5(6)-Carboxyrhodamine 6G-dUTP, 5-TAMRA-dUTP, and 5- ROX-dUTP.
  • Probes are either labeled by nick translation and PCR, or labeled by PCR using region specific primers, primers to vector/plasmid sequences, universal primers or degenerative oligonucleotide primers.
  • region specific primers primers to vector/plasmid sequences
  • primers to vector/plasmid sequences primers to vector/plasmid sequences
  • universal primers primers to degenerative oligonucleotide primers.
  • FISH Fluorescent In Situ Hybridization
  • a probe provided by the invention hybridizes (i.e., is complementary or substantially complementary) to a region of interest on a chromosome.
  • a probe can be designed to hybridize to any region on a chromosome.
  • a probe of the invention is used to detect the IGH locus.
  • the IGH locus includes variable (IGHV), diversity (IGHD), joining (IGHJ), and constant (IGHC) segments.
  • IGHC constant
  • a probe of the invention is used to detect IGHV and IGHC segments.
  • a probe of the invention may also be used to detect translocations involving the immunoglobulin heavy locus (IGH) on the long arm of chromosome 14 region 32.
  • IGH immunoglobulin heavy locus
  • translocations involving IGH on the long arm of chromosome 14 region 32 may be abbreviated as "14q32 translocations" or "t14q32".
  • 14q32 translocations involve many chromosomal partners (translocation partners).
  • 14q32 translocation partners in multiple myeloma may include, but are not limited to, Fibroblast Growth Factor Receptor 3 (FGFR3, human GenelD No. 2261 ), Wolf- Hirschhorn syndrome candidate 1 (WHSC1 or MMSET or WHSC1 /MMSET, human GenelD No.
  • Cyclin D3 (CCND3, human GenelD No. 896), Cyclin Dl (CCND1 , human GenelD No. 595), V-maf musculoaponeurotic fibrosarcoma oncogene homolog (MAF, human GenelD No. 4094), V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB, human GenelD No. 9935) and c-MYC (human GenelD No. 4609).
  • genomic locus refers to the specific location of a DNA sequence on a chromosome that encodes a polypeptide or RNA chain that has a function in the organism, and includes all associated regulatory regions, transcribed regions, termination sequences, and other functional sequence regions.
  • a probe of the invention is specific for a single genomic locus.
  • “specific” means a probe comprises a nucleic acid sequence that is complementary to a particular sequence of interest, e.g., a genomic locus or target transcript thereof.
  • a probe specific for a (e.g., single) genomic locus is not complementary or substantially complementary to, for example, a second genomic locus or a target transcript thereof.
  • the IGH locus is approximately 1 ,250 kb in length and comprises the four segments described above (i.e., IGHC, IGHJ, IGHD, IGHV), which encode an estimated 170-176 different genes. It has been discovered that (i) smaller probes to the IGH locus reduce the false positive rate of 14q32 translocations identified by current commercially available probes, and (ii) when a 14q32 translocation event occurs, IGHC drives expression of its translocation partner more efficiently than IGHV drives expression of its translocation partner.
  • a probe comprises a DNA sequence complementary to a chromosomal region comprising an entire genomic locus. In other embodiments, a probe comprises a DNA sequence complementary to a chromosomal region comprising a portion of a genomic locus. In still other embodiments, a probe comprises a DNA sequence complementary to a chromosomal region comprising some or all of a genomic locus, as well as DNA sequences upstream and/or downstream of the genomic locus. In exemplary embodiments, a probe comprises a DNA sequence specific for a single genomic locus. In other exemplary embodiments, a probe
  • a probe comprises a DNA sequence specific for segment of a genomic locus.
  • a probe comprises a DNA sequence complementary to a chromosomal region encoding a genomic locus comprising IGHC, IGHV, FGFR3, WHSC1 /MMSET, CCND1 , CCND3, MAF, MAFB, c-MYC, or a combination thereof.
  • a probe comprises a DNA sequence specific to a genomic locus comprising IGHC, IGHV, FGFR3, WHSC l/MMSET, CCND1 , CCND3, MAF, MAFB, c- MYC, or a combination thereof.
  • a probe comprises a DNA sequence complementary to a chromosomal region encoding a genomic locus comprising IGHC or IGHV. In another specific embodiment, a probe comprises a DNA sequence specific to a genomic locus comprising IGHC or IGHV.
  • a probe of the invention is about 1 to about 500 kilobases (kb) in length or about 50 to about 500 kb in length. In some embodiments, a probe is about 1 to about 100 kb in length. For example, a probe is about 1 to about 50 kb, or about 50 to about 100 kb in length. In other embodiments, a probe is about 100 to about 200 kb in length.
  • a probe is about 100 to about 150 kb, or about 150 to about 200 kb in length. In still other embodiments, a probe is about 200 to about 300 kb in length. For example, a probe is about 200 to about 250 kb, or about 250 to about 300 kb in length. In still other embodiments, a probe is about 300 to about 400 kb in length. For example, a probe is about 300 to about 350 kb, or about 350 to about 400 kb in length. In yet other embodiments, a probe is about 400 to about 500 kb in length. For example, a probe is about 400 to about 450 kb, or about 450 to about 500 kb in length. In some exemplary embodiments, a probe is about 100-300 kb in length. In other exemplary embodiments, a probe is about 200-400 kb in length. In still other exemplary
  • a probe is about 300-500 kb in length.
  • any suitable source (i.e., template) of nucleic acid may be used to produce a probe of the invention.
  • the source is a vector comprising the DNA sequence complementary to a region of interest on a chromosome.
  • vector refers to an autonomously replicating nucleic acid unit.
  • the present invention can be practiced with any known type of vector, including viral, cosmid, plasmid, artificial chromosomes, and plasmid vectors.
  • Non-limiting examples of artificial chromosomes include BACs (bacterial artificial chromosomes), PACs (PI
  • bacteriophage-derived artificial chromosome bacteriophage-derived artificial chromosome
  • YACs yeast artificial chromosomes
  • Alternative sources of DNA for FISH include, but are not limited to, genomic DNA from flow sorted chromosomes and genomic DNA from microdissected DNA.
  • the source of DNA for a probe is a BAC. In other exemplary
  • a source of DNA for a probe is a PAC.
  • a source of DNA for a probe is a YAC.
  • Vectors containing DNA complementary to a region of interest on a chromosome may be purchased through commercial vendors, or may be generated by the methods known in the art (Osoegawa K., et al, 2001 Genome Res 1 1 (3):483- 96).
  • a source of DNA for a probe is selected from the group of BAC clones listed in Table 1.
  • a source of DNA for a probe is selected from the IGHC and IGHV clones listed in Table 1.
  • a FISH probe of the invention may be used for cytogenetic and genomic analyses using methods well known in the art. Generally, locus specific probes are used to detect the desired loci, gene fusions, translocations, deletions, and amplifications. According to the invention, methods of the invention encompass means to detect IGHC and IGHV loci.
  • a method of the invention provides a means to detect IGHC and IGHV in a sample by metaphase FISH or interphase FISH using (at least) a pair of FISH probes.
  • the two probes are labeled with different fSuorochromes. each f!uorochrome emitting at different ultraviolet-wavelengths. If the two probes are brought into dose proximity, they may appear as two colors near each other, or the two colors may appear as a third color (for example, a red probe and green probe may produce a yellow signal). If the two probes are not brought Into close proximity, they may appear as two colors near each other, or they may appear as two colors not near each other.
  • a first probe comprises a DNA sequence complementary to a chromosomal region encoding a genomic locus or a segment of a genomic locus from IGH
  • a second probe comprises a nucleic acid sequence complementary to a chromosomal region encoding a genomic locus from IGHV.
  • a first probe comprises a nucleic acid sequence specific for a genomic locus or a segment of a genomic locus from IGHC
  • a second probe comprises a DNA specific for a genomic locus from IGHV
  • a first probe comprises a DNA sequence specific for a genomic locus or a segment of a genomic locus from IGHC
  • a second probe comprises a DNA sequence
  • a first probe comprises a DNA sequence complementary to a chromosomal region encoding a genomic locus or a segment of a genomic locus from the group consisting of IGH, and a second probe comprises a DNA sequence specific for a genomic locus from IGHV.
  • Substantially similar probes to any of the forgoing probes can also be used consonant with the invention.
  • Substantially similar probes are those that hybridize to the reference probes, or their complements, under highly stringent hybridization conditions or, in other embodiments, are at least about: 85, 70, 75, 80, 85, 90, 91 , 92, 93, 94, 95, 98, 97, 98, 99%, or more identical to the reference probe.
  • a first probe comprises a DNA sequence specific for IGHC
  • a second probe comprises a DNA sequence specific for IGHV
  • Suitable fiuorochromes and probes are further described in Section i.
  • the methods of the invention may be performed on any sample comprising a nucleic acid, for example, containing any cell type with a nucleus.
  • a cell may or may not have a 14q32 translocation.
  • a cell has a 14q23 translocation.
  • a cell does not have a 14q32 translocation.
  • a sample is comprised of cells grown in vitro. In other
  • a sample is a biological sample.
  • biological sample refers to a sample derived (i.e., isolated) from a subject.
  • a biological sample is modified, e.g., through staining or detectable labeling, such that the biological sample is not naturally occurring and is instead a product of human ingenuity.
  • Suitable subjects also referred to as individuals, may include a human, a livestock animal, a companion animal, a laboratory animal, or a zoological animal.
  • a subject may be a rodent, e.g., a mouse, a rat, a guinea pig, etc.
  • a subject may be a livestock animal.
  • Non-limiting examples of suitable livestock animals may include pigs, cows, horses, goats, sheep, llamas and alpacas.
  • a subject may be a companion animal.
  • companion animals may include pets such as dogs, cats, rabbits, and birds.
  • a subject may be a zoological animal.
  • a "zoological animal" refers to an animal that may be found in a zoo. Such animals may include non-human primates, large cats, wolves, and bears.
  • a subject may be a laboratory animal.
  • Non-limiting examples of a laboratory animal may include rodents, canines, felines, and non-human primates.
  • a subject is human.
  • the subject may or may not have been diagnosed with a disease or condition associated with a 14q32 translocation.
  • IGH translocations are common in B- cell malignancies.
  • B-cell malignancies include precursor B-cell acute lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-CLL), B-cell non-Hodgkin's lymphoma (including, but not limited to, follicular, mantle cell, Burkitt, diffuse large B-cell, marginal cell, mucosa-associated lymphoid tissue (MALT) lymphoma, and lymphoplasmacytic lymphoma) and myeloma.
  • B-ALL precursor B-cell acute lymphoblastic leukemia
  • B-CLL B-cell chronic lymphoblastic leukemia
  • B-cell non-Hodgkin's lymphoma including, but not limited to, follicular, mantle cell, Burkitt, diffuse large B-cell, marginal cell, mucos
  • the subject has clinical signs of disease. In yet other embodiments, the subject may be at risk for disease. In still other embodiments, the subject has been diagnosed with disease. In some exemplary embodiments, a subject has a tumor. In other exemplary embodiments, a subject has lymphoma. In still other exemplary embodiments, a subject has leukemia. In yet other exemplary embodiments, a subject has a B-cell malignancy. In a preferred embodiment, a subject has multiple myeloma.
  • Multiple myeloma includes symptomatic myeloma, asymptomatic myeloma, and monoclonal gammopathy of undetermined significance (MGUS), as defined in Kyle and Rajkumar Leukemia 23 :3-9 (2009, PubMedID 18971951 , incorporated by reference in its entirety), as well as the other stratifications and stages described in Kyle and Rajkumar 2009.
  • the method of the invention may be used with any of the numerous types of biological samples known in the art.
  • tissue samples or bodily fluids may include tissue samples or bodily fluids.
  • the biological sample is a tissue sample such as a tissue biopsy— e.g. a sample containing plasma cells, such as a bone marrow sample.
  • the tissue biopsy may be a biopsy of a tumor.
  • the biopsied tissue may be fixed, embedded in paraffin or plastic, and sectioned, or the biopsied tissue may be frozen and cryosectioned.
  • the biopsied tissue may be processed into individual cells or an explant, or processed into a homogenate, a cell extract, or a membranous fraction.
  • the sample may also be primary and/or transformed cell cultures derived from tissue from the subject.
  • the sample may be a bodily fluid.
  • bodily fluids include cerebrospinal fluid, interstitial fluid, blood, serum, plasma, saliva, sputum, semen, tears, lymph and urine.
  • the fluid may be used "as is", the cellular components may be isolated from the fluid, or a protein faction may be isolated from the fluid using standard techniques. In some preferred
  • the sample may be selected from the group consisting of blood, plasma, serum or lymph.
  • the sample is a biopsy of a tumor.
  • the sample is bone marrow.
  • the amount of bone marrow collected is 5 cc or less.
  • the amount of bone marrow collected is 5 cc, 4 cc, 3 cc, 2 cc or 1 cc.
  • the amount of bone marrow collected is 5 cc or less, 4 cc or less, 3 cc or less, 2 cc or less, or 1 cc or less.
  • the amount of bone marrow collected is 1 cc or less.
  • the method of collecting a biological sample can and will vary depending upon the nature of the biological sample and the type of analysis to be performed. Any of a variety of methods generally known in the art may be utilized to collect a biological sample. Generally speaking, the method preferably maintains the integrity of the sample such that it can be accurately measured according to the method of the invention.
  • metaphase FISH requires the availability of viable cells from which to produce chromosome preparations for analysis, whereas interphase FISH is able to make use of preserved cellular material and may thus be extended to include the use of paraffin embedded biopsy sections.
  • the collected sample may or may not be purified for plasma cells. Methods of purification biological samples for plasma cells are known in the art. In a specific embodiment, plasma cells are not purified from the biological sample. Instead FISH is performed on the sample without plasma cell purification.
  • the invention provides a method of developing a predictive model to determine the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in-situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone in the biological sample for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from the biological sample of each healthy individual is plotted as a point on an X/Y plot thereby generating
  • FISH flu
  • the invention provides a method to determine the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in-situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone in the biological sample for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone in the biological sample from each healthy individual is plotted as a point on an X/Y plot generating a normal clonal distribution
  • the invention provides a method of determining the treatment of a subject with pre-active multiple myeloma.
  • the method comprises: (a) obtaining biological samples from a cohort of healthy individuals; (b) performing fluorescence in-situ hybridization (FISH) on the biological samples from the cohort of healthy individuals using at least a first probe and a second probe, wherein the first probe comprises a nucleic acid sequence specific for IGHC and the second probe comprises a nucleic acid sequence specific for IGHV; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone for each healthy individual within the cohort of healthy individuals, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from each healthy individual is plotted as a point on an X/Y plot generating a normal clonal distribution; (d) calculating a region that encompasses the normal clonal distribution from the
  • the invention provides a method of determining the risk of a subject with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment.
  • the method comprises: (a) obtaining a biological sample from a subject diagnosed with pre-active multiple myeloma; (b) performing fluorescence in-situ hybridization (FISH) on the biological sample from the subject diagnosed with pre-active multiple myeloma using the same probes as used to develop the predictive model of the invention; (c) determining the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone from the subject diagnosed with pre-active multiple myeloma, wherein the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from the subject diagnosed with pre-active multiple myeloma is plotted as a point on the predictive model of the invention; and (d) determining the location of the plotted
  • Suitable biological samples and subjects are described in Section 11(a)
  • suitable probes are described in Section I
  • suitable methods of performing FISH are described in Section 11(a).
  • B-cell malignancies include precursor B-cell acute lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia (B-ALL), B-cell chronic lymphoblastic leukemia
  • B-CLL lymphoblastic leukemia
  • B-cell non-Hodgkin's lymphoma including, but not limited to, follicular, mantle cell, Burkitt, diffuse large B-cell, marginal cell, mucosa- associated lymphoid tissue (MALT) lymphoma, and lymphoplasmacytic lymphoma
  • myeloma myeloma
  • the cohort of healthy individuals is a group of individuals that have no signs or symptoms of multiple myeloma including, but not limited to, MGUS, asymptomatic multiple myeloma (AMM), smoldering multiple myeloma (SMM), indolent multiple myeloma, symptomatic MM and active multiple myeloma and have not been diagnosed with MGUS, AMM, SMM, indolent multiple myeloma, symptomatic MM or active MM. Since MGUS, AMM, SMM, indolent MM, symptomatic MM or active MM may or may not have symptoms, it is important that methodologies beyond symptomatology are used to define a cohort of healthy individuals.
  • a healthy individual may not be diagnosed with MGUS, SMM or MM according to the criteria presented in Table 1.
  • Table 1 For more information, see Kyle and Rajkumar, Leukemia 2009; 23(1 ): 3-9, which is hereby incorporated by reference in its entirety.
  • a subject with "pre-active multiple myeloma” is a patient diagnosed with MGUS, AMM, SMM, indolent MM, or any other pre-symptomatic or pre-active multiple myeloma condition.
  • a skilled artisan would be able to diagnose a subject with pre-active multiple myeloma.
  • the artisan may use the criteria described in Table 1 or the information provided in Kyle and Rajkumar, Leukemia 2009; 23(1 ): 3-9, which is hereby incorporated by reference in its entirety.
  • the pre-active multiple myeloma is selected from the group consisting of monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM) and asymptomatic multiple myeloma (AMM)
  • MGUS monoclonal gammopathy of undetermined significance
  • SMM smoldering multiple myeloma
  • ALM asymptomatic multiple myeloma
  • the cohort of healthy individuals may be 5 or more individuals.
  • the cohort of healthy individuals may be 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 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, or 50 individuals.
  • the cohort of healthy individuals may be more than 50 individuals.
  • the cohort of healthy individuals may be 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99 or 100 individuals.
  • the cohort of healthy individuals may be more than 100 individuals.
  • the cohort of healthy individuals may be about 100, about 1 10, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190 or about 200 individuals.
  • the cohort of health individuals may be more than 200 individuals.
  • FISH fluorescence in-situ hybridization
  • FISH is performed on biological samples from a cohort of healthy individuals.
  • FISH may be as described in Section ll(a).
  • FISH is performed with a first probe comprising a nucleic acid sequence specific for IGHC and the second probe comprising a nucleic acid sequence specific for IGHV.
  • the probes are labeled with two different fluorochromes such that they can be
  • first probe comprising a nucleic acid sequence specific for IGHC is labeled with a green fluorochrome and second probe comprising a nucleic acid sequence specific for IGHV is labeled with a red fluorochrome.
  • first probe comprising a nucleic acid sequence specific for IGHC is labeled with a red fluorochrome and second probe comprising a nucleic acid sequence specific for IGHV is labeled with a green fluorochrome.
  • the biological samples from a cohort of healthy individuals may be further stained to detect immunoglobulin light chains.
  • immunoglobulin light chains include Kappa light chain and Lambda light chain.
  • a biological sample may be stained to detect immunoglobulin Kappa light chain and immunoglobulin Lambda light chain in the same biological sample.
  • a biological sample may be separated into two aliquots and each aliquot may be individually stained to detect immunoglobulin Kappa light chain in one aliquot and immunoglobulin Lambda light chain in the other aliquot.
  • a biological sample is separated into two aliquots and each aliquot is individually stained to detect immunoglobulin Kappa light chain in one aliquot and immunoglobulin Lambda light chain in the other aliquot
  • a biological sample may be stained with an epitope-binding agent specific to an immunoglobulin light chain.
  • epitope-binding agent refers to an antibody, an aptamer, a nucleic acid, an oligonucleic acid, an amino acid, a peptide, a polypeptide, a protein, a lipid, a metabolite, a small molecule, or a fragment thereof that recognizes and is capable of binding to an immunoglobulin light chain.
  • a biological sample is stained with an antibody specific to an immunoglobulin light chain.
  • a biological sample is stained with an antibody specific to immunoglobulin Kappa light chain.
  • a biological sample is stained with an antibody specific to immunoglobulin Lambda light chain.
  • the antibody may be labeled with a fluorochrome. Suitable fluorochromes are described in Section 1(a). If the Kappa and Lambda light chains are to be detected in the same biological sample, then two different fluorochromes must be used. If the Kappa and Lambda light chains are to be detected in two aliquots from the same biological sample, then two different fluorochromes may be used or the same fluorochrome may be used. The fluorochrome used for the immunoglobulin light chains must be distinguishable from the fluorochromes used in FISH. In a specific embodiment, the fluorochrome used to detect immunoglobulin light chains is a blue fluorochrome.
  • the abundance of a dominant Kappa chain clone and the abundance of a dominant Lambda chain clone for each individual is determined.
  • a biological sample is examined. Any means to detect the fluorochromes may be used to examine the stained biological sample. In a specific embodiment, fluorescent
  • microscopy may be used to examine the stained biological sample.
  • each biological sample is examined for FISH staining and for immunoglobulin light chain staining.
  • the examination allows the determination of the abundance of the dominant clone for each immunoglobulin light chain. Accordingly, the abundance of a dominant clone for the Lambda light chain is determined and the abundance of a dominant clone for the Kappa light chain is determined.
  • the "dominant clone” refers to the clone that is most abundant in the biological sample.
  • Clones are cells with the same combination and pattern of fluorochrome staining. Stated another way, clones are cells with the same immunoglobulin light chain staining and the same pattern of FISH staining. For example, Kappa stained cells with the same combination and pattern of fluorochromes corresponding to the first probe and fluorochromes corresponding to the second probe are recognized as identical clones. Or, Lambda stained cells with the same combination and pattern of fluorochromes corresponding to the first probe and fluorochromes corresponding to the second probe are recognized as identical clones.
  • a Kappa or Lambda stained cell may have different patterns of FISH staining and each different pattern is a different clone.
  • To determine the amount of each clone the number of Kappa cells with the same pattern of FISH staining are counted and the number of Lambda cells with the same pattern of FISH staining are counted.
  • the amount of each clone is used to calculate the abundance of the clone.
  • the abundance of each clone is calculated as the amount of each clone divided by the total number of Kappa and Lambda stained cells in the biological sample. The value obtained is the abundance of that clone.
  • the value obtained for the most abundant clone among the Kappa cells and the value obtained for the most abundant clone among the Lambda cells is plotted as a point on an X/Y graph.
  • Lambda is plotted along the X-axis and Kappa is plotted along the Y-axis.
  • Kappa is plotted along the X-axis and Lambda is plotted along the Y-axis.
  • the determination of the abundance of a dominant Kappa clone and the abundance of a dominant Lambda clone is done for a biological sample from each individual in the cohort of healthy individuals.
  • the dominant Kappa clone and the dominant Lambda clone for each individual in the cohort of healthy individuals is plotted as a point on an X/Y plot generating a normal clonal distribution among healthy individuals.
  • a region encompassing the normal clonal distribution from the cohort of healthy individuals is calculated.
  • a prediction ellipse may be created based on the mean of all normal Lambda strata (x- axis) versus all normal Kappa strata (y-axis) with 99% confidence level.
  • the region is an ellipsoid.
  • the region may be calculated with greater than 85% confidence.
  • the region may be calculated with greater than 85%, greater than 90%, greater than 95%, or greater than 99% confidence.
  • the region may be calculated with greater than 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
  • the region may be calculated with 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
  • the calculated region may be calculated with 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
  • encompassing the normal clonal distribution from a cohort of healthy individuals generates a predictive model such that when a point is inside the region the risk of progressing to active multiple myeloma is low and when a point is outside the region the risk of progressing to active multiple myeloma requiring treatment is high.
  • the invention also encompasses a method of determining the risk of patients with pre-active multiple myeloma of progressing to active multiple myeloma requiring treatment. In another aspect, the invention also encompasses a method of determining the treatment of a subject with pre-active multiple myeloma.
  • a biological sample is obtained from a subject diagnosed with pre- active multiple myeloma.
  • the biological sample, subject, and pre-active multiple myeloma are as described above.
  • FISH is then performed as described above on the biological sample with the same probes as used to develop the predictive model.
  • the abundance of the dominant Kappa chain clone and the abundance of the dominant Lambda chain clone from the subject diagnosed with pre-active multiple myeloma is determined as described above and plotted as a point on the plot generated in the predictive model.
  • the location of the plotted point determines the risk of patients with pre-active multiple myeloma of progressing to symptomatic multiple myeloma requiring treatment. If the point is inside the region designated by the predictive model, then the subject is at low risk of progressing to active multiple myeloma. Alternatively, if the point is outside the region designated by the predictive model, then the subject is at high risk of progressing to active multiple myeloma and in need of treatment. A subject inside the region designated by the predictive model may be classified as having AMG
  • a subject outside the region designated by the predictive model may be classified as having PPCD (polyclonal plasma cell dyscrasias).
  • the subject has a high risk of progressing to active multiple myeloma.
  • the risk may be greater than about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • the risk may be greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • the risk may continue to increase over time. For example, the risk may be about 25% at one year, about 35% at two years, and about 65% at five years after initial cancer diagnosis.
  • the subject has a low risk of progressing to active multiple myeloma.
  • the risk may be less than about 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%.
  • the risk may be less than about 20%, 15%, 10%, or 5%.
  • the risk may be low, but may still increase over time. For example, the risk may be less than 10% at two years.
  • Increased or decreased "risk” or “probability” may be determined, for example, by comparison to the average risk or probability of an individual patient diagnosed with pre-active multiple myeloma of progressing to active multiple myeloma.
  • the overall proportion of patients who are diagnosed with active multiple myeloma within 5 years of diagnosis of pre-active multiple myeloma may be 50%.
  • an increased risk for an individual will mean that they are more than 50% likely to develop active multiple myeloma within 5 years, whereas a reduced risk will mean that they are less than 50% likely to develop active multiple myeloma.
  • Such comparisons may, in some circumstances, be made within patient populations limited or grouped using other factors such as age, ethnicity, serum albumin, and/or the presence or absence of other risk factors.
  • a subject may be predicted to be at high risk of progressing to active multiple myeloma requiring treatment. Accordingly, the methods disclosed herein may be used to select treatment for cancer patients. Specifically, the methods disclosed herein may be used to select treatment for pre-active multiple myeloma patients. In an embodiment, the subject is treated based on the outcome of the predictive model. As explained herein, the determination of a dominant Kappa clone and a dominant Lambda clone classify a subject as having a low or high risk of progressing to active multiple myeloma. This classification may be used to identify groups that are in need of treatment or not.
  • treatment means any treatment suitable for the treatment of B-cell malignancies.
  • B-cell malignancies may be treated with chemotherapy, radiotherapy, immunotherapy, and bone marrow transplant.
  • chemotherapy include proteosome inhibitors (e.g.
  • alkylating agents e.g., melphalan, cyclophosphamide, cisplatin, carboplatin, oxaliplatin
  • anti-metabolites paclitaxel, docetaxel
  • vinca alkaloids e.g.
  • the treatment is chemotherapy.
  • the treatment is radiotherapy.
  • the treatment is immunotherapy.
  • the treatment is bone marrow transplant.
  • the treatment is a proteosome inhibitor.
  • the treatment is bortezomib.
  • the methodology disclosed herein may be used to determine the change in risk of the subject over time.
  • the predictive model may be used to assess the risk of a subject at one point in time, then at a later time, the predictive model may be used to determine the change in risk of the subject over time.
  • the predictive model may be used on the same subject days, weeks, months or years following the initial determination of risk. Accordingly, the predictive model may be used to follow a subject with pre-active multiple myeloma to determine when the risk of progressing to active multiple myeloma is high thereby requiring treatment.
  • the methodology disclosed herein may also be used to determine the response to treatment.
  • patients who respond to treatment are said to have benefited from treatment.
  • Typical responses to treatment measured in clinical practice include, but are not limited to, overall survival, event free survival, time to progression, time to death, partial response, and complete response. These terms are well known in the art and are intended to refer to specific parameters measured during clinical trials and in clinical practice by a skilled artisan.
  • the predictive model may be performed on the subject prior to initiation of treatment, then at a later time, the predictive model may be used to determine the response to treatment over time.
  • the predictive model may be used on the same subject days, weeks, months or years following initiation of treatment.
  • the predictive model may be used to follow a subject receiving treatment to determine if the subject is responding to treatment. If the subject is inside the region defined by the predictive model, then the subject may be responding to treatment. If the subject is outside the region defined by the predictive model, then the subject may not be responding to treatment. These steps may be repeated to determine the response to therapy over time.
  • the methodology disclosed herein may also be used to identify relapse in a multiple myeloma subject who has achieved a complete response (CR).
  • the predictive model may be performed on the subject following determination of a CR, then at a later time, the predictive model may be used to determine the maintenance of a CR over time.
  • the predictive model may be used on the same subject days, weeks, months or years following determination of a CR.
  • the predictive model may be used to follow a subject achieving a CR to determine if the subject may relapse. If the subject is outside the region defined by the predictive model, then the subject is at risk for relapse and may require treatment. If the subject is inside the region defined by the predictive model, then the subject is at low risk for relapse and may not require treatment.
  • the invention also provides non-transient computer readable media comprising instructions that, when executed by a processor, cause the processor to perform steps consonant with the invention (e.g., the predictive model).
  • the instructions may entail accepting data representing the abundance of the dominant Kappa clone and the abundance of the dominant Lambda clone of the subject with pre-active multiple myeloma, comparing the values to predictive model (e.g. controls) and providing an evaluation of the data— e.g. whether the subject is inside or outside the region designated by the predictive model, and any implication (e.g. whether treatment should or should not be administered).
  • the invention also provides systems comprising these computer readable media and a processor adapted to execute the instructions.
  • Example 1 Defining Risk of MGUS and AMM Progression to Myeloma by Ig Heavy-chain FISH.
  • IGHC immunoglobulin heavy chain constant region
  • IGHV variable regions
  • Results IGHC and IGHV pattern diversity of PCs was present in all specimens, including healthy individuals. A bivariate distribution of polyclonal diversities among healthy individuals was calculated to create an ellipse defining the normal population pattern. Applying the ellipsoid definition to the patient populations, two subgroups could be distinguished in MGUS/AMM cases: one within the ellipse, representing patients with polyclonal PCs (without a dominant clone), while the other, who had a dominant clone with identical IGHC/ IGHV FISH pattern, fell outside of the ellipse and was termed asymptomatic monoclonal gammopathy (AMG; Dhodapkar et al.
  • AMG asymptomatic monoclonal gammopathy
  • the AMG subgroup among AMM patients was at high risk of progressing to symptomatic myeloma; one year progression to MM requiring therapy was 25%, 2-year progression was 38%, and the 5-year progression rate was estimated at 64% (FIG. 2).
  • the subgroup with polyclonal PCs in AMM had significantly lower risk of disease progression (p ⁇ 0.0001 ).
  • serum albumin ⁇ 3.5 g/dL
  • GEP70 risk score > -0.26
  • t(4;14) were significant progression factors of AMG patients (Hazard Ratios >2.2; p ⁇ 0.005).
  • age >65 yr, HR 2.05)
  • serum albumin ⁇ 3.5 g/dL, HR 3.21
  • having AMM HR 3.02

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Abstract

La présente invention concerne un modèle prédictif pour déterminer un traitement pour des sujets chez lesquels a été diagnostiqué un myélome multiple asymptomatique.
PCT/US2014/063719 2014-11-03 2014-11-03 Procédés de détection d'une chaîne lourde d'immunoglobuline et leurs procédés d'utilisation Ceased WO2016072969A1 (fr)

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