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WO2012142330A1 - Utilisation de micro-arn comme marqueurs de diagnostic et agents thérapeutiques dans le cancer de l'ovaire et les tumeurs métastasées disséminées dans la cavité péritonéale - Google Patents

Utilisation de micro-arn comme marqueurs de diagnostic et agents thérapeutiques dans le cancer de l'ovaire et les tumeurs métastasées disséminées dans la cavité péritonéale Download PDF

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Publication number
WO2012142330A1
WO2012142330A1 PCT/US2012/033386 US2012033386W WO2012142330A1 WO 2012142330 A1 WO2012142330 A1 WO 2012142330A1 US 2012033386 W US2012033386 W US 2012033386W WO 2012142330 A1 WO2012142330 A1 WO 2012142330A1
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mir
hsa
mrna
tumor
cancer
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Alexander S. BRODSKY
Laurent Brard
Hsin Ta WU
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Brown University
Women and Infants Hospital of Rhode Island
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Brown University
Women and Infants Hospital of Rhode Island
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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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • 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
    • C12Q2600/118—Prognosis of disease development
    • 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

Definitions

  • This invention relates generally to the field of cancer.
  • a biomarker is an identifying or distinguishing characteristic that is objectively measured and evaluated as an indicator of a normal biologic process, pathogenic process, or pharmacologic response to a therapeutic intervention.
  • a biomarker often refers to a substance or process that is indicative of the presence of cancer in the body.
  • a biomarker might be either a molecule secreted by a tumor or it can be a specific response of the body to the presence of cancer.
  • Genetic, epigenetic, proteomic, glycomic, and imaging biomarkers can be used for cancer diagnosis, prognosis and epidemiology.
  • Epithelial ovarian cancer is one of the most lethal gynecologic malignancy, affecting nearly twenty-two thousand women in the United States in 2010, and accounting for nearly fourteen thousand deaths during that year. Serous adenocarcinomas represent the majority of new EOC diagnoses, with most patients presenting symptoms during advanced stages of the disease. Despite removal of the ovaries, surgical debulking, and chemotherapy, most patients with serous EOC will suffer recurrences and ultimately succumb to the disease. Due to the limited efficacy of currently available treatment options for advanced EOC, there is a pressing need to develop new strategies to diagnose and treat ovarian cancer and other metastatic tumors. SUMMARY OF THE INVENTION
  • the invention represents a major advance in the diagnosis, prognosis, and treatment of tumors of the peritoneal cavity by providing micro-ribonucleic acids
  • microRNAs/miRNAs that are differentially expressed in primary and metastatic tumors (e.g. , serous epithelial ovarian cancer (EOC)), and which act as biomarkers to predict the severity of cancer and how patients respond to treatment.
  • EOC serous epithelial ovarian cancer
  • the compositions and methods of the invention predict time to disease progression and overall survival.
  • the subject is preferably a mammal in need of such treatment, e.g., a subject that has been diagnosed with a primary tumor in an organ or tissue of the peritoneal cavity.
  • the mammal can be, e.g. , any mammal, e.g. , a human, a primate, a mouse, a rat, a dog, a cat, a cow, a horse, or a pig.
  • the mammal is a human.
  • a composition for predicting presence of secondary site metastases or a predisposition thereto in a subject diagnosed as having a primary tumor comprises a detection reagent specific for at least one microRNA sequence selected from the group consisting of hsa-miR-146a, hsa-miR-150, hsa-miR-193a-5p, hsa-miR-31, hsa-miR-21, hsa-miR-370, hsa- let-7d, hsa-miR-29a, hsa-miR-508-5p, hsa-miR-152, hsa-miR-509-3-5p, hsa-miR-508-3p, hsa-miR-708, hsa-miR-214, hsa-miR-431, hsa-miR-185, hsa-miR-124,
  • the composition further comprises a second detection reagent specific for at least one mRNA selected from the group consisting of INTS4, NARS2, SNORD31, INTS2, TRIP10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1, ITGA11, IGHM, COL8A1, NNMT, COL1A1, BGN, INHBA, and COL11 Al (Group I).
  • a second detection reagent specific for at least one mRNA selected from the group consisting of INTS4, NARS2, SNORD31, INTS2, TRIP10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1, ITGA11, IGHM, COL8A1, NNMT, COL1A1, BGN, INHBA, and COL11 Al (Group I).
  • the composition comprises a detection reagent specific for at least one mRNA selected from the group consisting of FNTS4, NARS2, SNORD31, INTS2, TRIP10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1, ITGA11, IGHM, COL8A1, NNMT, COL1A1, BGN, FNHBA, and COL11A1 (i.e., in the absence of a detection reagent specific for a microRNA described above).
  • a detection reagent specific for at least one mRNA selected from the group consisting of FNTS4, NARS2, SNORD31, INTS2, TRIP10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1, ITGA11, IGHM, COL8A1, NNMT, COL
  • the composition optionally comprises a second detection reagent specific for at least one mRNA selected from the group consisting of COPZ2, NUCB1, LPL, CCDC49, GFPT2, LOX, NNMT, RGS1, ASNA1, FXYD5, SERPINE1, KIF26B, S100A10, ALDH1A3, CALB2, and PLAUR (Group II).
  • a second detection reagent specific for at least one mRNA selected from the group consisting of COPZ2, NUCB1, LPL, CCDC49, GFPT2, LOX, NNMT, RGS1, ASNA1, FXYD5, SERPINE1, KIF26B, S100A10, ALDH1A3, CALB2, and PLAUR (Group II).
  • compositions for miRNA detection/quantification include those in which the miRNA is selected from the group consisting of hsa-let-7d, hsa-miR-146a, hsa-miR-29a, hsa-miR-193a-5p, hsa-miR-31, hsa-miR-21, hsa-miR-708, hsa-miR-152, hsa-miR-214, and hsa-miR-150.
  • the miRNA is selected from the group consisting of hsa- let-7d, hsa-miR-146a, hsa-miR-29a, hsa-miR-193a-5p, hsa-miR-31, and hsa-miR-150.
  • the miRNA is selected from the group consisting of hsa-miR-146a, hsa- miR-193a-5p, hsa-miR-31, and hss-miR-150.
  • the microRNA comprises a combination of two microRNAS selected from the combinations listed in Table 10, a combination of three microRNAS selected from the combinations listed in Table 11, or a combination of four microRNAS selected from the combinations listed in Table 12.
  • mRNAs are useful as independent prognostic tools and together with miRNAs offer improved prognostic capability.
  • detection/quantifiction of microRNAs is combined with detection/quantification of one or more mRNAs selected from the group consisting of mRNAs listed in Table 1 A or IB.
  • a method for predicting the presence of secondary site metastases or a predisposition thereto in a subject diagnosed as having a primary tumor is carried out by providing a tissue sample obtained from the primary tumor; detecting in the tissue sample at least two biomarkers selected from the group consisting of hsa-miR-146a, hsa-miR-150, hsa-miR- 193a-5p, hsa-miR-31, hsa-miR-21, hsa-miR-370, hsa-let-7d, hsa-miR-29a, hsa-miR-508-5p, hsa-miR-152, hsa-miR-509-3-5p, hsa-miR-508-3p, hsa-miR-708, hsa-miR-214, hsa-miR-431, hsa-miR-185,
  • the data is calculated, correlated, and compared, e.g., using a computer, to yield a prognosis for metastases or survival.
  • the method optionally further comprises detecting in the tissue sample a messenger ribonucleic acid (mRNA) transcript encoding INTS4, NARS2, SNORD31, INTS2, TRIP 10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1, ITGA1 1, IGHM, COL8A1, NNMT, COL1A1, BGN, INHBA, and
  • mRNA messenger ribonucleic acid
  • a lower level of the mRNA transcript compared to the control level of the mRNA indicates that the subject is suffering from or at an increased risk of developing secondary site metastases.
  • compositions and methods of identifying and inhibiting or mimicking (or augmenting/replacing) microR As that promote the
  • the primary tumor is a tumor located within the peritoneal cavity, and the metastasis is intra-abdominal metastases or lymph node metastases.
  • the primary tumor comprises serous, clear cell, endometrioid, or mucinous carcinoma cells.
  • compositions and methods described herein are useful in the diagnosis, prognosis, and treatment of various primary tumors and metastases, e.g., epithelial ovarian cancer (EOC), pancreatic cancer, gastric, kidney, colorectal cancer, hepatic cancer, bladder cancer, or breast cancer.
  • EOC epithelial ovarian cancer
  • pancreatic cancer gastric, kidney, colorectal cancer, hepatic cancer, bladder cancer, or breast cancer.
  • the tumor is within the peritoneal cavity or the abdominal-pelvic cavity.
  • the primary tumor comprises serous epithelial ovarian cancer (EOC), the most aggressive and lethal form of EOC.
  • the invention provides methods for predicting an increased risk of developing distant metastatic neoplastic disease ⁇ i.e., secondary site metastases) in a subject diagnosed as comprising a primary tumor.
  • increased risk is meant a risk greater than that typically associated with individuals comprising a primary tumor.
  • the level of at least one biomarker e.g., at least one, at least two, at least three, at least four, or at least five biomarkers
  • a tissue sample obtained from a primary tumor, tissue/organ from which the tumor was derived, or bodily fluid from the affected subject, e.g., a subject diagnosed as having an ovarian tumor or a lump that is diagnosed as being suspect of a malignant condition.
  • the tissue sample is obtained from a bodily fluid, e.g., blood, serum, lymphatic fluid, plasma, urine, saliva, semen, or breast milk from a subject.
  • a difference in level miR or mRNA levels comprises at least 10%, 20%, 50%>, 2-fold, 10- fold, 20-fold or more compared to a control or reference value.
  • a control or reference value comprises an average of measurements of at least 100 primary ovarian tumors.
  • mir-146a and mir-150 are separate from the average by 50% on the Agilent arrays in the TCGA data, and there is a two-fold difference in expression for mir- 146a and mir-150 in the high and low risk groups.
  • the invention also encompasses testing expression of the above-described microRNAs and mRNAs in metastatic tumors using the same methods.
  • the change in expression of the microRNA or mRNA between the primary tumor and metastatic tumor is measured and quantified.
  • the difference in expression level is indicative of patient outcome, i.e., a greater difference corresponds to poor prognosis. If upregulation of a given microRNA or mRNA is correlated with a higher risk of developing metastatic disease, an even higher level of expression of the microRNA or mRNA in a sample of a metastatic tumor from the same individual indicates a poor outcome (more severe disease) in that individual, and vice versa.
  • miRNA expression signatures i.e., biomarkers
  • biomarkers are selected from the group consisting of micro ribonucleic acid-31 (miR-31 ; GenBank Accession Number NR 029505.1
  • miR-124 GenBank Accession Number NR 029670.1 (GL262205258), NR 029669.1 (GL262205253), or NR 029668.1
  • miR-152 GenBank Accession Number NR 029687.1 (GL262205334), incorporated herein by reference
  • miR-146a GenBank Accession Number NR 029701.1 (GL262205399), incorporated herein by reference
  • miR-431 Gene ID: 574038, incorporated herein by reference
  • miR-214 GeneBank Accession Number NR 029627.1 (GL262206305), incorporated herein by reference.
  • the miRNA biomarkers are selected from the group consisting of hsa-miR-193a-5p, hsa-miR-708, hsa- miR-31, hsa-miR-508-3p, hsa-miR-124, and hsa-miR-185.
  • the at least two biomarkers are detected via quantitative real time polymerase chain reaction (qPCR).
  • qPCR quantitative real time polymerase chain reaction
  • tissue/tumor sample is compared to a control level obtained from a normal tissue.
  • control level is meant a level previously determined to be associated with non-metastatic tumors.
  • the control level is obtained from a normal tissue sample.
  • normal tissue is meant non-cancerous tissue.
  • An increase in the level of at least two biomarkers in the tumor sample indicates that the subject is suffering from or at an increased risk of developing a malignant tumor at an anatomical site distant from the primary tumor, thereby predicting an increased risk of developing distant metastatic neoplastic disease.
  • a distant metastasis from ovarian cancer is a tumor that occurs in an organ or tissue other than ovarian tissue.
  • the tumor is an endocrine tumor such as breast cancer, ovarian cancer, colon cancer, prostate cancer and endometrial cancer.
  • metastases to the lymph nodes are typically used for initial staging; however, the methods of the invention are also useful to predict lymph node involvement such as axillary lymph node involvement. The methods are also useful to predict time course of disease progression and to predict patient survival.
  • a messenger ribonucleic acid (mRNA) transcript is optionally detected in the tissue sample.
  • methods for predicting an increased risk of developing distant metastatic neoplastic disease (i.e., secondary site metastases) in a subject diagnosed as comprising a primary tumor include detecting an mRNA transcript in a tissue sample without prior identification of miRNA biomarkers, as described above.
  • a tissue sample is obtained from a primary tumor, tissue/organ from which the tumor was derived, or bodily fluid from the affected subject, e.g., a subject diagnosed as having an ovarian tumor or a lump that is diagnosed as being suspect of a malignant condition.
  • the tissue sample is obtained from a bodily fluid, e.g., blood, serum, lymphatic fluid, plasma, urine, saliva, semen, or breast milk from a subject.
  • a bodily fluid e.g., blood, serum, lymphatic fluid, plasma, urine, saliva, semen, or breast milk from a subject.
  • at least one mRNA transcript is detected, e.g., at least two, at least three, at least four, or at least five mRNA transcripts are detected.
  • the mRNA transcript is detected via quantitative real time reverse transcription polymerase polymerase chain reaction (qRT-PCR).
  • qRT-PCR quantitative real time reverse transcription polymerase polymerase chain reaction
  • Suitable mRNA transcripts include those encoding programmed cell death protein 4 (PDCD4; GenBank Accession Number CAI40095.1 (GI:57162254), incorporated herein by reference), serine/threonine-protein kinase 38-like (STK38L; GenBank Accession Number NP_055815.1 (GL24307971), incorporated herein by reference), reversion-inducing-cysteine-rich protein with kazal motifs (RECK; GenBank Accession Number AAH60806.1 (GL38174247), incorporated herein by reference), ELAV-like protein 1 (HuR; GenBank Accession Number NP 001410.2
  • NUMB protein numb homolog
  • GenBank Accession Number AAD01548.1 GL4102705
  • 14-3-3 protein epsilon YWHAE
  • GenBank Accession Number NP 006752.1 GL5803225
  • AT-rich interactive domain-containing protein 1 A ARID 1 A
  • GenBank Accession Number AC049597.1 GL226346315
  • the level of the at least one mRNA transcript in the tissue/tumor sample is compared to a control level obtained from a normal tissue.
  • a control level is obtained from a non-cancerous tissue sample.
  • a decrease (i.e., downregulation) in the level of mRNA in the tumor sample compared to the control level indicates that the subject is suffering from or at an increased risk of developing distant metastatic neoplastic disease.
  • an increase (i.e., upregulation) in the level of mRNA in the tumor sample compared to the control level indicates that the subject is suffering from or at an increased risk of developing distant metastatic neoplastic disease.
  • the primary tumor comprises serous, clear cell, endometrioid, or mucinous carcinoma cells.
  • the tumor is within the peritoneal cavity.
  • EOC epithelial ovarian cancer
  • pancreatic cancer colorectal cancer
  • hepatic cancer hepatic cancer
  • bladder cancer hepatic cancer
  • breast cancer hepatic cancer
  • Also provided are methods of treating a peritoneal cavity tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a composition comprising an inhibitor of a miRNA selected from the group consisting of miR-31, miR-21, miR-124, miR-150, miR-185, miR-708, miR-193-5p, miR-29a, let-7d, miR- 886-3p, miR-270, miR-152, miR-146a, miR-431, and miR-214.
  • Suitable inhibitors include anti-miRs, antagomiRs, peptide nucleic acid (PNA) locked nucleic acid (LNA), or small molecule inhibitors.
  • Also described herein are methods of treating a peritoneal cavity tumor in a subject in need thereof comprising administering to a subject a therapeutically effective amount of a composition comprising miR-508-3p, miR-509-3-5p, or miR-508-5p.
  • synthetic miRNA mimics that have the same sequence as the depleted, naturally occurring miRNA are administered.
  • Suitable miRNA mimics include a miR-509-3-5p mimic, a miR-508-3p mimic, and a miR-508-5p mimic.
  • Also provided is a method of inhibiting cancer metastases in the peritoneal cavity of a subject comprising modulating the level of miR-31, miR-21, miR-124, miR-150, miR-185, miR-708, miR-193-5p, miR-29a, let-7d, miR-886-3p, miR-270, miR-152, miR-146a, miR-431, miR-214, miR-508-3p, miR-509-3-5p, or miR-508-5p to change mRNA transcript levels.
  • the mRNA encodes PDCD4, STK38L, RECK, HuR, NUMB, YWHAE, or ARID 1 A.
  • the level of mRNA and encoded protein would also be modulated. In this manner, proteins are utilized as markers to evaluate the therapeutic efficacy of miRNA administration.
  • a method of prognosis for ovarian cancer patients includes the steps of detecting various miRNAs (described above) in a sample of ovarian tissue or bodily fluid of the patient following excision of a primary tumor, wherein an elevation in the level of various miRNA compared to a normal control level or over time indicates recurrence of malignancy.
  • a method for predicting survival time of a cancer patient is carried out by detecting various miRNAs (described above) in a tissue biopsy in which an increase in various miRNAs levels is correlated with a decrease in survival time.
  • the assay for prediction of survival of the individual or recurrence of a tumor is carried out before or after any treatment of the patient for the cancer.
  • Purified defines a degree of sterility that is safe for administration to a human subject, e.g., lacking infectious or toxic agents.
  • an "isolated” or “purified” nucleic acid molecule, polynucleotide, polypeptide, or protein is substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized.
  • Purified compounds are at least 60% by weight (dry weight) the compound of interest.
  • the preparation is at least 75%, more preferably at least 90%>, and most preferably at least 99%>, by weight the compound of interest.
  • RNA Ribonucleic acid
  • DNA deoxyribonucleic acid
  • substantially pure is meant a nucleic acid, polypeptide, or other molecule that has been separated from the components that naturally accompany it.
  • the polynucleotide, polypeptide, or other molecule is substantially pure when it is at least 60%, 70%, 80%, 90%, 95%, or even 99%, by weight, free from the proteins and naturally-occurring organic molecules with which it is naturally associated.
  • a substantially pure polypeptide may be obtained by extraction from a natural source, by expression of a recombinant nucleic acid in a cell that does not normally express that protein, or by chemical synthesis.
  • a substantially pure nucleic acid ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) is free of the genes or sequences that flank it in its naturally- occurring state.
  • an “effective amount” and “therapeutically effective amount” of a formulation or formulation component is meant a nontoxic but sufficient amount of the formulation or component to provide the desired effect.
  • an “effective amount” is meant an amount of a compound, alone or in a combination, required to reduce or prevent the growth or invasiveness of a tumor of the peritoneal cavity in a mammal.
  • the tumor is of an organ of the female reproductive system such as a breast or an ovarian tumor.
  • the effective amount of active compound(s) varies depending upon the route of administration, age, body weight, and general health of the subject. Ultimately, the attending physician or veterinarian will decide the appropriate amount and dosage regimen.
  • the methods are also useful to inform choice of therapy, because gene expression signatures and levels of expression indicate whether or not the patient being treated will respond to Platinum/taxane therapy.
  • the chemotherapy drugs used to treat ovarian cancer are fairly standard. Typically doctors combine a platinum-based drug such as carboplatin
  • a method of determining whether a tumor is resistant to a chemotherapeutic agent is carried out by detecting in a tissue sample from a subject at least one biomarker selected from the group consisting of hsa-miR-146a, hsa-miR-150, hsa-miR-193a-5p, hsa- miR-31, hsa-miR-21, hsa-miR-370, hsa-let-7d, hsa-miR-29a, hsa-miR-508-5p, hsa-miR-152, hsa-miR-509-3-5p, hsa-miR-508-3p, hsa-miR-708, hsa-mi
  • mRNA markers are also suitable for this purpose used either alone or in combination with an evaluation of microRNAs.
  • a method of determining whether a tumor is resistant to a chemotherapeutic agent is carried out by detecting in a tissue sample from a subject a messenger ribonucleic acid (mRNA) transcript encoding at least one of INTS4, NARS2, SNORD31, INTS2, TRIP10, RASSF2, GADD45B, LTBP2, MYH9, MMP14, PLAU, OLFML2B, THY1, CRISPLD2, COMP, FNDC1 , ITGA11, IGHM, COL8A1, NNMT, COL1A1, BGN, INHBA, and COL11A1 or a mRNA transcript in Table 1 or Table 2 and comparing the levels of the mRNA transcript in the tissue sample to a control level of the mRNA, wherein a lower level of the INTS4, NARS2, SNORD31, INTS2 mRNA transcript compared to the control
  • encoding at least one of COPZ2, NUCB1, LPL, CCDC49, GFPT2, LOX, NNMT, RGS1, ASNA1, FXYD5, SERPINE1, KIF26B, S100A10, ALDH1A3, CALB2, and PLAUR and comparing the levels of the mRNA transcript in a tissue sample to a control level of the mRNA, wherein a higher level of mRNA indicates that subject comprises a drug resistant tumor.
  • the information generated by these methods is useful to a physician. If the microRNA and/or mRNA profile indicates that the tumor(s) are drug resistant to platinum and/or taxane drugs, the physician would choose an alternative therapeutic strategy.
  • Figure 1 is a schematic showing that metastatic tumors are clonal expansions with phenotypic diversity.
  • Figure 1 A is a schematic showing hierarchical clustering of array comparative genomic hybridization (aCGH) data for primary and metastatic tumors. The results show that each matched pair of primary (P) and omental metastases (M) are more similar to each other than tumors from other patients. Segments were mapped to genes before clustering.
  • Figure IB is a schematic showing hierarchical clustering of micro- ribonucleic acids (microRNAs/miRNAs). The results suggests that more primary and metastatic pairs are not alike, which implies phenotypic diversity.
  • microRNAs/miRNAs micro- ribonucleic acids
  • Figure 2 is a series of graphs and photomicrographs showing that metastases exhibit lower expression of cell cycle checkpoints and are more proliferative compared to primary tumors.
  • Figure 2A is a Gene Set Enrichment Analysis (GSEA) curve indicating that G2/M cell cycle checkpoints are exhibit higher expression in primary tumors compared to metastases.
  • Figure 2B is a photomicrograph demonstrating that metastases have higher Ki-67 proliferation indexes than primary tumors, which is consistent with cell cycle checkpoint expression. Blue indicates hematoxylin and eosin (H&E) staining of tumor cells consistent with lower expression of cell cycle checkpoints in metastases. Brown indicates
  • IHC immunohistochemistry
  • Figure 3 is a series of bar charts and photomicrographs showing that miR-21 is overexpressed in metastases compared to primary tumors.
  • Figure 3 A is a bar chart showing the results of a top 10 (p ⁇ 0.03) Taqman qPCR screen from bulk tumor.
  • Figure 3B is a bar chart showing the results of RNA Taqman qPCR on tumor cells isolated by Laser Capture Microdissection (LCM).
  • Figure 3C is a series of photomicrographs of in situ hybridization (ISH) of two cases illustrating increased expression of miR-21 in metastases.
  • Blue staining is ISH signal. Red staining is nuclear Red staining of tumor cells. Arrows indicate Blue staining of miR-21 signal. Increased miR-21 signal originates from tumor cells and not just stroma.
  • Figure 4 is a series of photomicrographs and bar charts demonstrating that metastatic miRNAs promote anchorage independent growth in ovarian cancer cells.
  • Figure 4C shows that inhibition of miR-21 and miR-31 by peptide nucleic acids (PNAs) reduces spheroid size in soft agar in ovarian carcinoma cells (OVCAR-8).
  • PNAs peptide nucleic acids
  • Figure 4D shows that that inhibition of miR-21 and miR-31 by peptide nucleic acids (PNAs) reduces the number of colonies in soft agar in OVCAR-8 cells. Error bars are standard deviation. P-values were calculated by Student's t-test. miR-31 did not significantly affect ES-2 cells in either spheroid or soft agar assays.
  • Figure 4E is a bar chart showing that free miRNA expression levels in OVCAR-8 spheroids were reduced after inhibition, as determined by Taqman after non-organic RNA purification preserving the miRNA-PNA interaction.
  • Figure 5 is a schematic and series of line graphs showing that a metastasis expression signature (both mRNAs and miRNAs) predicts survival. The number of patients in each curve is noted.
  • Figure 5 A is a flow-chart of classification using Cox univariate analysis, support vector machine (SVM) analysis, and Kaplan-Meier analysis.
  • Figure 5B is a line graph showing that the 32 gene metastasis signature distinguishes patients by their overall survival. The lower curve is strongly biased towards higher expression of up-regulated metastasis genes.
  • Figure 5C is a line graph showing that out of the top 10 differentially expressed miRNAs, 6 contribute significantly to Kaplan-Meier analysis of survival including miR-31. miR-21 was relatively uniformly expressed in primary tumors in this dataset.
  • miRNA classification was performed using a combination SVM and logistic model with leave one out validation.
  • Figure 6 is a schematic showing the experimental approach for sampling tumors. Specimens are sampled from each quadrant and compared to each other.
  • Figure 7 is a series of graphs and a chart demonstrating that miR-21 and mir-31 mRNA-predicted TargetScan targets are globally repressed in metastases compared to primary tumors.
  • Figure 7A is a line graph showing global distribution of the Spearman correlation coefficients between mRNA targets and miR-31 and miR-21. miRNA mRNA targets predicted by PITA (red line) are TargetScan (blue line) are enriched for negatively correlated transcripts consistent with being down-regulated compared to randomly selected sets of transcripts permuted 1,000 times (grey lines).
  • Figure 7B is a dot plot showing miR-21 and the literature validated miR-21 target, programmed cell death protein 4 (PDCD4), are negatively linearly correlated.
  • PDCD4 programmed cell death protein 4
  • Figure 7C is a chart showing that genes with Spearman coefficients ⁇ -0.5 are significantly enriched for specific pathways and functions as determined by IPA. P-values are multiple hypothesis corrected using Benjamini-Hochberg. Selected genes for each pathway are listed, thereby representing high quality candidates.
  • Figure 8 is a series of line graphs, bar graphs, and photomicrographs showing that metastases are more proliferative and include less apoptotic cells than matched primary tumors.
  • Figure 8 A is a line graphs showing a GSEA enrichment plot that suggests Reactome G2/M cell cycle checkpoints including Chekl, CCNB2, and BUB1 are expressed higher in primary tumors.
  • Figure 8B is a bar graph showing a qPCR validation of cell cycle checkpoints.
  • Figure 8C is a series of photomicrographs showing representative
  • FIG. 8D is a line graph showing a GSEA enrichment plot that suggests that repressors of apoptosis from Gene Ontology are expressed higher in metastases than primary tumors.
  • Figure 8E is a bar graph showing qPCR validation of selected genes.
  • Figure 8F is a series of photomicrographs showing that representative TUNEL staining reveals more apoptotic cells in primary tumors compared their matched omental metastases.
  • Figure 9 is a series of line graphs showing Kaplan-Meier analysis of metastasis gene expression signature (Group II list of genes). Black dotted lines indicates high risk group and the grey line indicates the low risk group.
  • Figure 9 A is a line graph showing the top 100 up- regulated genes in metastases applied to the TCGA Affymetrix dataset.
  • Figure 9B is a line graph showing the 32 genes significantly up-regulated in omental metastases and with HR>1, p ⁇ 0.05 by a Cox proportional hazards model in the TCGA Affymetrix U133 Plus 2.0 microarray dataset.
  • Figure 9C is a line graph showing the 28 genes significantly up-regulated in omental metastases and with HR>1, p ⁇ 0.05 in the TCGA Agilent 244K expression dataset.
  • Figure 9F is a line graph showing the 16 overlap genes applied to the Tothill data (Tothill RW, et al. (2008) Clin Cancer Res 14 (16):5198-5208).
  • Figure 9G-9I are a series of line graphs showing the combination of residual disease after debulking leads to robust discrimination, even for patients with sub-optimal debulking in Figure 91, on the TCGA Affymetrix U133 Plus 2.0 platform.
  • Figure 10 is a series of charts showing hierarchical clustering of copy number aberrations and mRNA expression.
  • Figure 10A is a chart showing that most patients cluster by their copy number aberrations (CNAs). The circles indicate matched pairs of primary and metastatic tumors that cluster together with similar patterns of CNAs. Clustering by mRNA expression levels in primary and metastatic tumors reveals a less ordered linkage.
  • Figure 1 OB is a series of representative heat maps of the copy number data from two cases. Many CNAs are found in both primary and metastatic tumors.
  • Figure 11 is a series of photomicrographs showing that tumors are of ovarian origin and are serous epithelial as indicated from examination of H&E and cytokeratin staining. Fiogure 11 shows H&E staining of two representative cases. CA125 and cytokeratin staining of one case is consistent with ovarian tumor origins.
  • Figure 12 is a series of bar charts showing microarray and qPCR measurements generally agree.
  • Figure 12 shows qPCR validation of genes primary and metastatic tumors from two cases.
  • Figure 13 is a series of line graphs and charts showing that enriched pathways reveal common features of metastases.
  • GSEA enrichment plot suggests that WNT/p-Catenin signaling from the Signaling Transduction database and double strand break repair gene ontology are up-regulated in metastases compared to primary tumors.
  • Figure 14 is a series of heat maps showing the distribution of expression levels in all TCGA patients ordered by survival. Green indicates higher expression and the gene expression signature is expressed higher in shorter survival patients.
  • Figure 15 is a series of line graphs showing Kaplan-Meier analysis of the 16 gene high-risk signature (Group II list of genes) applied to the TCGA Agilent microarray and Tothill datasets in combination with residual disease. Black dotted line indicates high risk group and grey line indicates low risk group.
  • Figure 16 is a schematic showing a network analysis of the 16 gene expression signature (Group II list of genes). 11 of the 16 genes are included in the network and are shaded in gray. Central regulatory nodes include NFKB, AKT, PDGF, and ⁇ -estradiol.
  • Figure 17 is a schematic showing a summary of all CNAs in all 12 cases. Most of the genome is subject to copy number aberrations consistent with extensive genomic instability of serous ovarian cancer. Green is amplification, red is copy loss. These data pertain to the Group II list of genes/gene expression signature.
  • Figure 18 is a schematic showing top IPA identified network in the 467genes up- regulated in metastases.
  • Pathway analysis identified increased TFGB1 signaling in metastases using Ingenuity software analysis.
  • Figure 19 is a line graph demonstrating the proportion survival of patients from 0 to 120 months after diagnosis. (Table 1A list of genes)
  • Figure 20 is a schematic demonstrating that 24 mR As significantly differentially upregulated in primary versus omental metastatic tumors distinguish primary and metastatic tumors.
  • Figure 21 is a series of graphs showing that a combination of mRNAs and miRNAs improve patient survival prognostic predictions. (Group I list of genes).
  • Figure 22 is a series of graphs showing that a combination of mRNAs and miRNAs improve patient survival prognostic predictions. (Group I list of genes).
  • Figure 23 is a series of graphs showing that a combination of mRNAs and 4 miRNAs improve patient survival prognostic predictions. (Group I list of genes).
  • Figure 24 is a series of graphs showing that an mRNA and miRNA signature combined with residual disease clear stratifies in high and low risk groups. (Group I list of genes).
  • Figure 25 is a series of line graphs demonstrating that miR-150 induces resistance and stimulates growth in SKOV-3 and IGROV-1 cells (miR-150 and miR-146a were key contributors to the survival curves described above). (Group I list of genes).
  • Epithelial ovarian cancer (EOC) is a deadly disease affecting thousands of women each year. Serous adenocarcinomas represent the majority of new EOC diagnoses, with most present in advanced stages. Most are treated by removal of the ovaries and debulking surgery followed by platinum-taxane-based therapy, with 5-year survival under 50%, as patents often suffer from extensive metastatic disease. While the 5 year survival rates are 94% for localized tumors, the survival rates drop to 28% after distant metastasis. Only 15% of cases are diagnosed while the cancer is still confined to the primary site, and 62% of ovarian cancers are not detected until after distant metastasis.
  • Ovarian cancer is a lethal disease, typically diagnosed in late stages. To treat ovarian cancer, it is commonly thought that early diagnosis through imaging and/or the development of a screening test could be a panacea (Moore RG, et al. Am J Obstet Gynecol.
  • Ovarian cancer likely has a different etiology, and appears to be particularly heterogeneous at the mRNA expression and DNA copy levels (Gorringe KL Mol Oncol. 2009;3(2): 157-64). Intra-tumor hetero eneity, sampling and metastasis
  • Intra-tumor heterogeneity provides a possible bottleneck in understanding and characterization of tumors, especially with gene expression where cell mixtures complicates interpretation (Marusyk A and Polyak K Biochim Biophys Acta. 2010;1805(1): 105-17.
  • PMCID 2814927. Genetic divergence between primary and metastases has been observed by monitoring DNA copy number (Navin N, et al. Genome Res. 2009) and DNA sequencing identified mutations (Yachida S, et al. Nature. 2010;467(7319): 1114-7). These data are consistent with metastases originating from rare tumor cells within a primary tumor and clonally expanding into more genetically homogeneous metastases. However, prior to the invention described herein, why particular regions of a tumor are more likely to metastasize, and if metastases are always more homogeneous was unclear (Marusyk A and Polyak K. Biochim Biophys Acta. 2010;1805(1): 105-17. PMCID: 2814927).
  • microRNAs as regulators of metastasis
  • MicroRNAs are small, noncoding single-stranded RNAs that comprise a class of gene regulators (Bader, et al. Cancer Research. 2010;70(18):7027-30, incorporated herein by reference). They are highly conserved from plants to humans and are encoded by their respective genes. miRNAs are transcribed from the genome as longer precursor molecules that are cleaved by the nuclear ribonuclease Drosha into approximately 70- to 100- nt-long oligonucleotides that form a distinct hairpin structure.
  • RNAse Dicer RNA-induced silencing complex
  • RISC RNA-induced silencing complex
  • RISC loaded with miRNA and the target mRNA inhibits the translation of the mRNA by either a silencing mechanismor by degradation of the mRNA.
  • the miRNA and mRNA sequences are merely partially complementary, which enables miRNAs to target a broad, but nevertheless a specific, set of mRNAs.
  • miRNAs that affect metastasis in model systems have differential expression in primary and metastatic tumors, further justifying the approach described herein (Valastyan S, et al. Cell. 2009;137(6): 1032-46).
  • miRNAs regulate gene expression post-transcriptionally affecting both mRNA stability and translation often repressing gene expression by 3 ' UTR binding (Filipowicz W, et al. Nat Rev Genet. 2008;9(2): 102-14).
  • miRNAs are critical regulators of metastasis in a number of cancers (Hurst DR, et al. Cancer Res.
  • miRNAs may be pleiotropic and affect multiple steps of the metastatic process (Valastyan S, et al. Cell. 2009;137(6): 1032-46; Valastyan S, et al. Genes Dev. 2009; Valastyan S, et al. Cancer Res. 2010;70(12):5147-54. PMCID: 2891350).
  • determining which mRNAs are targeted by miRNAs was difficult, as the many computational methods have very high false positive rates in any given condition as recently reviewed, even with genome -wide expression data (Thomas M, et al. Nat Struct Mol Biol. 2010; 17( 10): 1169-74).
  • miRNAs have been linked to metastasis, including miR-31 (Valastyan S, et al. Cell. 2009; 137(6): 1032-46; Dykxhoorn DM Cancer Res.
  • a qualitative analysis of the miRNAs (differentially) expressed in a particular sample is performed using known methods, e.g., the Agilent miRNA microarray platform (Agilent Technologies, Santa Clara, Calif, USA) according to the manufacturer's instructions.
  • Agilent miRNA microarray platform Agilent Technologies, Santa Clara, Calif, USA
  • Quantitative analysis (verification) of the miRNA expression data obtained is typically performed via real-time quantitative RT-PCR employing a TaqMan MicroRNA assay (Applied Biosystems, Foster City, Calif, USA) according to the manufacturer's instructions.
  • the quantification of the miRNAs may be performed by using real-time quantitative RT-PCR employing SYBR Green I (Sigma Aldrich Corporation, St. Louis, Mo., USA), an asymmetrical cyanine dye binding to double-stranded DNA.
  • SBS Illumina's sequencing by synthesis
  • HiSeq, TruSeq, MiSeq Life Technologies SOLiD Sequencing products are also useful detection/quantification.
  • primers are used to amplify the microRNA or mRNA target and process it into cDNA.
  • Such primers are at least 10 nucleotides in length (typically 17-25 nucleotides in length) and are complementary to the miRNA or mRNA sought to be evaluated.
  • Primer design to amplify a known sequence is well known in the art. All of these methods and devices are suitable for analysis of both microRNAs and mRNAs.
  • Kits or systems for evaluating miRNAs and/or mRNAs include a) a oligonucleotide complementary to an miRNA; and b) optionally, reagents for the formation of the
  • an apparatus or composition for diagnosing or prognosing metastases or survival in a patient with a cancer comprises a solid support, in which a surface of the solid support is linked to an
  • oligonucleotide complementary to an miRNA The level of an miRNA in a sample can be measured using any technique that is suitable for detecting RNA expression levels in a biological sample. Suitable techniques for determining RNA expression levels in cells from a biological sample are well known in the art. Examples of such techniques include, but are not limited to, Northern blot analysis, RT-PCR, microarrays, in situ hybridization. In a particular embodiment, a high-throughput system, for example, a microarray, is used to measure the expression level of a plurality of genes.
  • the plurality of nucleic acid molecules encoding a miRNA sequence that are comprised in a diagnostic kit of the present invention may include one or more "sense nucleic acid molecules" and/or one or more "anti-sense nucleic acid molecules".
  • the diagnostic kit includes one or more "sense nucleic acid molecules” (i.e. the miRNA sequences as such).
  • a diagnostic kit includes one or more "anti-sense nucleic acid molecules" (i.e.
  • the molecules may comprise probe molecules (for performing hybridization assays) and/or oligonucleotide primers (e.g., for reverse transcription or PCR applications) that are suitable for detecting and/or quantifying one or more particular (complementary) miRNA sequences in a given sample.
  • the relative number of miRNA gene transcripts in cells can also be determined by reverse transcription of miRNA gene transcripts, followed by amplification of the reverse- transcribed transcripts by polymerase chain reaction (RT-PCR), i.e., the miRNA is processed into cDNA and then analyzed.
  • RT-PCR polymerase chain reaction
  • the levels of miRNA gene transcripts can be quantified in comparison with an internal standard, for example, the level of mRNA from a
  • a suitable "housekeeping” gene for use as an internal standard includes, e.g., myosin or glyceraldehyde-3 -phosphate dehydrogenase (G3PDH).
  • G3PDH glyceraldehyde-3 -phosphate dehydrogenase
  • the methods for quantitative RT-PCR and variations thereof are within the skill in the art.
  • a high throughput stem loop real-time quantitative polymerase chain reaction (RT-qPCR) is used to detect miRNA expression.
  • a microchip i.e., a microarray
  • a microchip may be constructed containing a set of probe oligodeoxynucleotides that are specific for a set of miRNA genes.
  • the expression level of or or more miRNAs in a biological sample is determined by reverse transcribing the RNAs to generate a set of target oligodeoxynucleotides, and hybridizing them to probe oligodeoxynucleotides on the microarray to generate a hybridization, or expression, profile.
  • the hybridization profile of the test sample is then compared to the pre-determined expression level of a control sample to determine which miRNAs have an altered expression level in cancer cells.
  • probe oligonucleotide or “probe oligodeoxynucleotide” refers to an oligonucleotide that is capable of hybridizing to a target oligonucleotide.
  • Target oligonucleotide or “target oligodeoxynucleotide” refers to a molecule to be detected (e.g., via hybridization).
  • miRNA-specific probe oligonucleotide or “probe oligonucleotide specific for an miRNA” is meant a probe oligonucleotide that has a sequence selected to hybridize to a specific miRNA gene product, or to a reverse transcript of the specific miRNA gene product.
  • an "expression profile” or “hybridization profile” is essentially a fingerprint of the state of the sample; while two states may have any particular gene similarly expressed, the evaluation of a number of genes simultaneously allows the generation of a gene expression profile that is unique to the state of the cell. That is, normal tissue may be distinguished from a cancer tissue, and within a cancer tissue, different prognosis states (good or poor long term survival prospects, for example) may be determined. By comparing expression profiles of a cancer tissue in different states, information regarding which genes are important (including both up- and down-regulation of genes) in each of these states is obtained.
  • sequences that are differentially expressed in a cancer tissue or normal tissue allows the use of this information in a number of ways. For example, a particular treatment regime may be evaluated (e.g., to determine whether a chemotherapeutic drug act to improve the long-term prognosis in a particular patient). Similarly, diagnosis may be done or confirmed by comparing patient samples with the known expression profiles. Furthermore, these gene expression profiles (or individual genes) allow screening of drug candidates that suppress the cancer expression profile or convert a poor prognosis profile to a better prognosis profile.
  • the microarray can be prepared from gene-specific oligonucleotide probes generated from known miR A sequences.
  • the array contains one, two, three, four or more different oligonucleotide probes for each miRNA, one containing the active, mature sequence and the other being specific for the precursor of the miRNA.
  • the array may also contain controls, such as one or more mouse sequences differing from human orthologs by only a few bases, which can serve as controls for hybridization stringency conditions.
  • tRNAs from both species may also be printed on the microchip, providing an internal, relatively stable, positive control for specific hybridization.
  • One or more appropriate controls for nonspecific hybridization may also be included on the microchip. For this purpose, sequences are selected based upon the absence of any homology with any known miRNAs.
  • a microarray may be fabricated using techniques known in the art and printed using commercially available microarray systems, e.g., the GENEMACHINE, OMNIGRID 100 MICRO ARRAYER and AMERSHAM CODELINK activated slides.
  • Labeled cDNA oligomer corresponding to the target RNAs is prepared by reverse transcribing the target RNA with labeled primer. Following first strand synthesis, the RNA/DNA hybrids are denatured to degrade the RNA templates. The labeled target cDNAs thus prepared are then hybridized to the microarray chip under hybridizing conditions. At positions on the array where the immobilized probe DNA recognizes a complementary target cDNA in the sample, hybridization occurs.
  • the labeled target cDNA marks the exact position on the array where binding occurs, allowing automatic detection and quantification.
  • the output consists of a list of hybridization events, indicating the relative abundance of specific cDNA sequences, and therefore the relative abundance of the corresponding complementary miRNA, in the patient sample.
  • the labeled cDNA oligomer is a biotin-labeled cDNA, prepared from a biotin-labeled primer.
  • the microarray is then processed by direct detection of the biotin- containing transcripts using, e.g., streptavidin-alexa647 conjugate, and scanned utilizing conventional scanning methods. Image intensities of each spot on the array are proportional to the abundance of the corresponding miRNA in the patient sample.
  • primers for detection of miRNAs or mRNAs, primers (Taqman MicroRNA reagents, e.g., primer pairs, panels, cards are available from Life Technologies/ Applied Biosystems (Carlsbad, California). Other reagents for detection of microRNAs, e.g., miScript miRNA PCR Array, miScript II RT Kit) are available from Qiagen (Valencia, California). Assay kits for measuring microRNA expression are also available from Nano String (Seattle, WA), e.g., nCounter miRNA Expression Assay Kits. miRCURY LNATM Universal RT microRNA PCR is available from Exiqon (Vedbaek Denmark; Woburn, MA).
  • miRNAs miRNAs, mRNAs, and ovarian cancer
  • mRNA expression signatures can characterize different subtypes and predict prognosis of ovarian cancer (Tothill RW, et al. Clin Cancer Res. 2008;14(16):5198-208). miRNA expression signatures in metastasis and survival have not been investigated. The data presented herein is also applicable to large primary tumor studies such as TCGA to develop a possible prognosis expression signature. The integrated genomics approach of the national TCGA project identifies many mutations and candidate factors from primary tumors; however, prior to the invention described herein, their role in metastasis was unclear.
  • miR-21 usually promotes aggressive tumors, and is currently being targeted for therapeutic development (Medina PP, et al. Nature. 2010;467(7311):86-90; Connolly EC, et al. Mol Cancer Res. 2010;8(5):691-700; Bonci D. Recent Pat Cardiovasc Drug Discov.
  • miR-21 has the potential to regulate hundreds of transcripts, which suggests miR-21 acts through a range of mechanisms. Described herein are results that indicate that some "literature-validated" targets are likely being repressed by miR-21 during ovarian metastasis, while others are not. On the other hand, miR-31 inhibits breast cancer progression, and appears to act as a suppressor of metastasis (Valastyan S, et al. Cell.
  • miR-31 has higher expression in metastatic tumors demonstrating that it promotes metastasis.
  • the in vitro data described herein indicate that both miR-21 and miR-31 promote anchorage independent growth, consistent with the clinical observations.
  • miRNA have also been identified as targets for therapeutic intervention (Bader, et al. Cancer Research. 2010;70(18):7027-30, incorporated herein by reference).
  • the rationale for developing miRNA therapeutics is based on the premise that aberrantly expressed miRNAs play key roles in the development of human disease, and that correcting these miRNA deficiencies by either antagonizing or restoring miRNA function may provide a therapeutic benefit.
  • synthetic miRNA mimics that have the same sequence as the depleted, naturally occurring miRNA are administered to restore miRNA function.
  • miRNA function is inhibited by, e.g., anti-miRs, antagomiRs, peptide nucleic acid (PNA) locked nucleic acid (LNA), or small molecule inhibitors.
  • PNA peptide nucleic acid
  • LNA locked nucleic acid
  • mRNA expression signatures have been investigated to define subtypes and predict prognosis of ovarian cancer patients (Tothill RW, et al. Clin Cancer Res. 2008;14(16):5198- 208). Most of these signatures are developed by identifying a set of miRNAs or mRNAs that correlates with survival or some other clinical attribute. Sometimes mechanisms can be found suggesting that these factors are important. This approach often runs into statistical bias problems (Bair E and Tibshirani R. PLoS Biol. 2004;2(4):E108. PMCID: 387275; Shi L, et al. Nat Biotechnol. 2010;28(8):827-38; Majewski IJ and Bernards R. Nat Med.
  • miRNAs provide a practical tool to investigate metastasis, as they are relatively straightforward to measure and genetically manipulate.
  • the results described herein identify miR-21 and miR-31 as critical regulators of ovarian cancer metastasis. Similar to ovarian tumors, a number of common cancers metastasize to the omentum and within the abdominopelvic cavity including pancreatic, colo-rectal, and liver cancers. Thus, the results described below are also relevant general features important for dissemination within the abdominopelvic cavity (Lengyel E. Am J Pathol. 2010;177(3): 1053-64. PMCID: 2928939). Identifying key features of advanced disease also helps discriminate the most aggressive early lesions as early detection strategies improve.
  • EOC epithelial ovarian cancer
  • Described herein is a quadrant sampling approach to refine the characterization of ovarian tumors to gain insight into intra-tumor heterogeneity and to identify differentially expressed miRNAs regulating metastasis.
  • the often large serous ovarian tumors allow for extensive intra-tumor sampling, while the large dynamic range of Taqman qPCR is utilized to accurately and specifically identify miRNAs with expression differences between primary tumors and metastases.
  • the results are validated using in situ hybridization and other qPCR methods to gain high confidence in the genomic screening data.
  • the function and expression of miRNAs and mRNA targets in a panel of ovarian cancer cell lines to reduce the dependence on any one genetic background.
  • miR-21 and miR-31 are important to reduce apoptosis and to promote proliferation, respectively, leading to the more aggressive nature of cancer cells in metastases.
  • the data presented herein indicate an interesting role of these miRNAs in driving 3D spheroid formation and growth, but not proliferation in 2D, adherent culture.
  • the function of miR-21 and miR-31 in metastasis is determined, and mRNA targets of miR-21 and miR-31 that drive metastasis are identified.
  • the utility of 2D vs. 3D culture conditions for ovarian cancer cells is characterized by linking laboratory and clinical observations, and an approach is established to link human tumor observations with mechanisms and regulation of metastasis in xenograft mouse models.
  • miRNA expression signatures i.e., biomarkers
  • miR-31 and miR-21 are examined herein because both have been previously associated with cancer, and because their changing expression levels was validated by multiple assays, as described below. Indeed, miR-21 and miR-31 are expressed higher in metastases in 89% and 78% of patients, respectively.
  • miR-31 is the top-ranked miRNA in the primary vs. metastases comparison, and miR-31 is significantly associated with more aggressive disease in a miRNA expression signature.
  • miR-21 is one of the first miRNAs identified to promote growth and robustness of cancer cells, but has not been extensively examined in ovarian cancer.
  • miRNAs e.g. , miR-31 and miR-21
  • miR-21 are metastatic regulators.
  • described herein is the role of miRNA in promoting metastasis.
  • miR-150 is intergenic on chromosome 19 and thus is not obviously co-regulated with gene. miR-150 regulates B-cell differentiation and the timing of expression is critical for its proper role in promoting B cell development. Prior to the invention, there was little known about the role of miR-150 in cancer. Previous reports suggested that miR-150 can either promote or inhibit tumors, highlighting the common theme of context dependent functions of miRNAs. Using primary/metastatic tumor data, the results described herein does not indicate an inverse correlation with the expression of previously identified miR-150 targets P2RX7 or EGR2.
  • mRNA expression signatures i.e., biomarkers
  • mRNA transcripts that are upregulated between primary and metastatic tumors include those set forth in Table 1.
  • EPYC NM_004950 (NM_004950.4 GL223941903)
  • TIMP3 NM_000362 (NM_000362.4 GL75905820) FGF7 NM_002009 (NM_002009.3 GL219842354)
  • CYP1B1 NM_000104 (NM_000104.3 GI: 189491762) PLAU NM_002658 (NM_002658.3 GL222537757) CDCP1 NM_022842 (NM_022842.3 GL30410804) TLR3 NM_003265 (NM_003265.2 GL 19718735) SVEP1 NM_153366 (NMJ53366.3 GL 148886653) MFAP4 NM_002404 (NM_002404.2 GI:310923209) Gene S ⁇ in hoi en liank Accession N umber
  • KGFLP1 NR_003674 (NR_003674.2 GL383276551)
  • FCGR2C NM_201563 (NM_201563.4 GL226874954)
  • KGFLP1 NR_003674 (NR_003674.2 GL383276551) LRP1 NM_002332 (NM_002332.2 GI: 126012561) HLA-F NM_001098479 (NM_001098479.1 GL 149158701) ALCAM NM_001627 (NM_001627.3 GL343168768) Gene S ⁇ in hoi en liank Accession N umber
  • ARPC1B NM_005720 (NM_005720.3 GL325197176)
  • CTGF NM_001901 (NM_001901.2 GL98986335)
  • FCGR1A NM_000566 (NM_000566.3 GL 167621452)
  • FCGR2A NM_001136219 (NM_001136219.1 GL210031821)
  • NUCB1 NM_006184 (NM_006184.5 GL297374833) Gene S ⁇ in hoi ( ien liank Accession N umber
  • VDR NM_001017535 (NM_001017535.1 GL63054844)
  • FCGR1B NM_001017986 (NM_001017986.3 GL349732137)
  • CTLA4 005214 (NM_ 005214.4 GI: :339276048)
  • FCGR1A 000566 (NM_ _000566.3 GI: : 167621452)
  • GPNMB NM_001005340 (NM_001005340.1 GL52694751)
  • ANXA2P2 // ANXA2P2 // ANXA2P2 NR_003573 (NR_003573.1 GL 148833516)
  • NM_182978 (NM_182978.2 GL215276940)
  • PTGFRN NM_020440 (NM_020440.2 GL41152505) Gene S ⁇ in hoi en liank Accession N umber
  • GBP1 NM_002053 (NM_002053.2 GL 166706902)
  • GK3P NR_026575 (NR_026575.1 GL219803770)
  • GIMAP8 NM_175571 (NMJ75571.2 GL55953077) Gene S ⁇ in hoi ( ien liank Accession N umber
  • IGHV4-31 AK301335 (AK301335.1 GL194376247)
  • Table IB Reduced list of upregulated mRNAs Gene Symbol mRNA Accession
  • Table 1C shows Group II mRNA expression signature: COPZ2, NUCBl, LPL, CCDC49, GFPT2, LOX, NNMT, RGSl, ASNAl, FXYD5, SERPP E1, KIF26B, SIOOAIO, ALDH1A3, CALB2, and PLAUR.
  • CALB2 NM_001740 calbindin 2 transcript variant CALB2
  • mRNAs listed above e.g., those listed in Table IB, distinguish primary and metastatic tumors and have prognostic significance on their own. When combined with the miRNA list, particularly strong patient stratification was observed.
  • Additional exemplary upregulated mRNA transcripts from Table 1 include FAM38B, COLEC12, GFPT2, LOX, KIF26B, CALB2, RGS4, FSTL3, PDGFA, KRT5, PTGIS, RGS1, SERPINE1, NUCB1, ADAM 12, MMP16, LPL, NNMT, ASNA1, APBB1IP, FXYD5, S100A10, ALDH1A3, CD1E, ZFHX4, C10orf26, CCDC49, EMR2, FAS, ERBB2, PLAUR, and CLASS.
  • Suitable mRNA transcripts that are downregulated between primary and metastatic tumors include those set forth in Table 2.
  • EFTUD1 NM_024580 (NM_024580.5 GI: 111120335)
  • TOM1L1 AB065085 (AB065085.1 GI:21104503) STAR NM_000349 (NM_000349.2 GL56243550)
  • PACRGL NMJ45048 (NM_145048.3 GL 195539373)
  • ANKRD36 AK304740 (AK304740.1 GL194388231)
  • NIPBL NM_015384 (NM_015384.4 GL 189163520)
  • CTDSPL2 NM_016396 (NM_016396.2 GL 100815974) ARHGAP1 IB NM_001039841 (NM_001039841.1 GI:89886349)
  • TAFIA NM_005681 (NM_001039841.1 GL89886349)
  • EIF2A NM_032025 (NM_032025.3 GL83656780)
  • TMEM128 NM_032927 NM_032927.2 GL39725660
  • TIPARP NM_015508 (NM_015508.4 GL296080689)
  • 0SGEPL1 NM_022353 (NM_022353.2 GI: 116812635) (icue S ⁇ mbol GenBank Accessions Number wnregulated III RNA) (incorporated herein by reference)
  • TRMT61B NM_017910 (NM_017910.3 GL222831586) MRS2 NM_020662 (NM_020662.2 GL93204868) CEP57 NM_014679 (NM_014679.4 GL344925821) Gene Symbol Gen Bank Accession Number (Downregulated mRNA) (incorporated herein by reference)

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Abstract

Cette invention concerne des compositions et des méthodes permettant de prévoir des métastases dans un cancer.
PCT/US2012/033386 2011-04-15 2012-04-12 Utilisation de micro-arn comme marqueurs de diagnostic et agents thérapeutiques dans le cancer de l'ovaire et les tumeurs métastasées disséminées dans la cavité péritonéale Ceased WO2012142330A1 (fr)

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013021088A3 (fr) * 2011-08-09 2013-08-01 Oncomatrix, S.L. Procédés et produits permettant le diagnostic in vitro, le pronostic in vitro et le développement de produits pharmaceutiques contre les carcinomes invasifs
WO2015049282A1 (fr) * 2013-10-02 2015-04-09 Oncomatrix, S.L. Procédés et produits pour le diagnostic et le pronostic d'une malignité tumorale ovarienne
WO2015128671A1 (fr) * 2014-02-27 2015-09-03 Queen Mary University Of London Marqueurs biologiques de l'endométriose
WO2017158358A1 (fr) * 2016-03-15 2017-09-21 Almac Diagnostics Limited Signatures géniques pour la détection et le traitement du cancer
WO2017170334A1 (fr) * 2016-03-28 2017-10-05 東レ株式会社 Composition pharmaceutique pour le traitement et/ou la prévention du cancer
WO2017214952A1 (fr) * 2016-06-16 2017-12-21 毛侃琅 Construction et application d'un vecteur lentiviral pour l'inhibition spécifique de l'expression de l'arnmi-185 humain
WO2017219166A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition simultanée de l'expression de deux miarn, et application associée
WO2017219165A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition spécifique de l'expression de miarn-29a humain et de mir-140 humain, et application associée
WO2017219169A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition de l'expression de miarn-29a et de mir-185, et son application
KR20180063004A (ko) * 2018-05-28 2018-06-11 의료법인 성광의료재단 Ｆｏｘａ２를 포함하는 난소암의 진단을 위한 마커 및 조성물
CN108410983A (zh) * 2018-02-11 2018-08-17 中山大学 Nkx2-8在肿瘤耐药检测中的应用及fao抑制剂在nkx2-8缺失型肿瘤中的应用
CN108586622A (zh) * 2018-05-11 2018-09-28 山东大学 Tat-pdcd4融合蛋白及其在治疗卵巢癌药物中的应用
CN110089081A (zh) * 2016-12-16 2019-08-02 松下知识产权经营株式会社 编码方法、解码方法、传输方法、解码设备、编码设备、传输设备
CN111041095A (zh) * 2019-07-11 2020-04-21 江苏医药职业学院 检测8号染色体开放阅读框73表达水平的试剂的应用和试剂盒
KR20220009332A (ko) * 2020-07-15 2022-01-24 국민대학교산학협력단 miR-214-3p를 포함하는 난소암에 대한 화학요법제의 저항성 예측용 조성물
US11371099B2 (en) 2015-11-30 2022-06-28 Mayo Foundation For Medical Education And Research HEATR1 as a marker for chemoresistance
CN115494237A (zh) * 2022-09-19 2022-12-20 浙江省人民医院 Rps6ka2在制备卵巢癌诊断试剂和治疗药物中的应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070212721A1 (en) * 2006-01-27 2007-09-13 Tripath Imaging, Inc. Methods for identifying patients with an increased likelihood of having ovarian cancer and compositions therefor
US20100240126A1 (en) * 2008-05-07 2010-09-23 Shi-Lung Lin Development of universal cancer drugs and vaccines
US20110033875A1 (en) * 2007-12-12 2011-02-10 University Of Georgia Research Foundation, Inc. Glycoprotein cancer biomarker

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070212721A1 (en) * 2006-01-27 2007-09-13 Tripath Imaging, Inc. Methods for identifying patients with an increased likelihood of having ovarian cancer and compositions therefor
US20110033875A1 (en) * 2007-12-12 2011-02-10 University Of Georgia Research Foundation, Inc. Glycoprotein cancer biomarker
US20100240126A1 (en) * 2008-05-07 2010-09-23 Shi-Lung Lin Development of universal cancer drugs and vaccines

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ASANGANI I.A. ET AL.: "MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer.", ONCOGENE, vol. 27, 2008, pages 2128 - 36, XP055073318, DOI: doi:10.1038/sj.onc.1210856 *
KOROBKO I.V.: "Gen proteinkinazy MAK-V i ryad drugikh genov s izmenennoi ekspressiei v opukholyakh i ikh ispolzovanie v onkologii.", AFTOREF. DISS. DOKTORA BIOL. NAUK. M., 2009, pages 47, 20, Retrieved from the Internet <URL:http://www.genebiology.ru/dissertation/korobko.pdf> [retrieved on 20120806] *
MITCHELL P.S.: "Circulating microRNAs as stable blood-based markers for cancer detection.", PNAS, vol. 105, no. 30, 29 July 2008 (2008-07-29), pages 10513 - 10518 *
YAN L-X. ET AL.: "MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph nodemetastasis and patient poor prognosis.", RNA, vol. 14, 2008, pages 2348, Retrieved from the Internet <URL:http://rnajournal.cshlp.org/content/14/11/2348.short> [retrieved on 20120821] *

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WO2013021088A3 (fr) * 2011-08-09 2013-08-01 Oncomatrix, S.L. Procédés et produits permettant le diagnostic in vitro, le pronostic in vitro et le développement de produits pharmaceutiques contre les carcinomes invasifs
US9702879B2 (en) 2011-08-09 2017-07-11 Oncomatryx Biopharma, S.L. Methods and products for in vitro diagnosis, in vitro prognosis and the development of drugs against invasive carcinomas
WO2015049282A1 (fr) * 2013-10-02 2015-04-09 Oncomatrix, S.L. Procédés et produits pour le diagnostic et le pronostic d'une malignité tumorale ovarienne
WO2015128671A1 (fr) * 2014-02-27 2015-09-03 Queen Mary University Of London Marqueurs biologiques de l'endométriose
JP2017509333A (ja) * 2014-02-27 2017-04-06 クイーン マリー ユニバーシティ オブ ロンドン 子宮内膜症に対するバイオマーカー
US11371099B2 (en) 2015-11-30 2022-06-28 Mayo Foundation For Medical Education And Research HEATR1 as a marker for chemoresistance
WO2017158358A1 (fr) * 2016-03-15 2017-09-21 Almac Diagnostics Limited Signatures géniques pour la détection et le traitement du cancer
JPWO2017170334A1 (ja) * 2016-03-28 2019-02-07 東レ株式会社 癌の治療及び／又は予防用医薬組成物
WO2017170334A1 (fr) * 2016-03-28 2017-10-05 東レ株式会社 Composition pharmaceutique pour le traitement et/ou la prévention du cancer
CN108883176A (zh) * 2016-03-28 2018-11-23 东丽株式会社 癌的治疗和/或预防用药物组合物
RU2746123C2 (ru) * 2016-03-28 2021-04-07 Торэй Индастриз, Инк. Фармацевтическая композиция для лечения и/или предотвращения злокачественных опухолей
WO2017214952A1 (fr) * 2016-06-16 2017-12-21 毛侃琅 Construction et application d'un vecteur lentiviral pour l'inhibition spécifique de l'expression de l'arnmi-185 humain
WO2017219165A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition spécifique de l'expression de miarn-29a humain et de mir-140 humain, et application associée
WO2017219169A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition de l'expression de miarn-29a et de mir-185, et son application
WO2017219166A1 (fr) * 2016-06-19 2017-12-28 毛侃琅 Vecteur lentiviral pour l'inhibition simultanée de l'expression de deux miarn, et application associée
CN110089081A (zh) * 2016-12-16 2019-08-02 松下知识产权经营株式会社 编码方法、解码方法、传输方法、解码设备、编码设备、传输设备
CN108410983A (zh) * 2018-02-11 2018-08-17 中山大学 Nkx2-8在肿瘤耐药检测中的应用及fao抑制剂在nkx2-8缺失型肿瘤中的应用
CN108410983B (zh) * 2018-02-11 2021-08-17 中山大学 Nkx2-8在肿瘤耐药检测中的应用及fao抑制剂在nkx2-8缺失型肿瘤中的应用
CN108586622A (zh) * 2018-05-11 2018-09-28 山东大学 Tat-pdcd4融合蛋白及其在治疗卵巢癌药物中的应用
KR101890387B1 (ko) 2018-05-28 2018-08-21 의료법인 성광의료재단 Ｆｏｘａ２를 포함하는 난소암의 진단을 위한 마커 및 조성물
KR20180063004A (ko) * 2018-05-28 2018-06-11 의료법인 성광의료재단 Ｆｏｘａ２를 포함하는 난소암의 진단을 위한 마커 및 조성물
CN111041095A (zh) * 2019-07-11 2020-04-21 江苏医药职业学院 检测8号染色体开放阅读框73表达水平的试剂的应用和试剂盒
CN111041095B (zh) * 2019-07-11 2021-11-30 江苏医药职业学院 检测8号染色体开放阅读框73表达水平的试剂的应用和试剂盒
KR20220009332A (ko) * 2020-07-15 2022-01-24 국민대학교산학협력단 miR-214-3p를 포함하는 난소암에 대한 화학요법제의 저항성 예측용 조성물
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CN115494237A (zh) * 2022-09-19 2022-12-20 浙江省人民医院 Rps6ka2在制备卵巢癌诊断试剂和治疗药物中的应用

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