Materials and Methods Related to Modulation of Mismatch Repair and Genomic Stability by miR-155

Abstract

The present invention provides materials and methods related to modulation of mismatch and genomic stability by miR-155.

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         5 . A method of identifying mismatch repair (MMR) dysfunctional cells in a test sample, comprising
 comparing miR-155 levels in a test sample to miR-155 levels of a control sample, wherein differentially-expressed miR-155 levels identify the test sample, relative to the level of the corresponding miR-155 in the control sample, as containing MMR dysfunctional cells.   
     
     
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         10 . A method of providing a prognosis in a patient, comprising:
 comparing miR-155 levels in a test sample from the patient to miR-155 levels in a control sample, wherein downregulated miR-155 levels indicates level of the miR-155 in the test sample, relative to the level of the corresponding miR-155 in the control sample, provides the patient with a poor prognosis.   
     
     
         11 . A method of treating mismatch repair (MMR) dysfunction in patient in need of such treatment, comprising administering to the patient a pharmaceutically-effective amount of a composition of anti-sense miR-155. 
     
     
         12 . A method of  claim 11 , wherein the MMR dysfunction is selected from the group consisting of: neuroblastoma; lung cancer; bile duct cancer; non small cell lung carcinoma; hepatocellular carcinoma; lymphoma; nasopharyngeal carcinoma; ovarian cancer; head and neck squamous cell carcinoma; squamous cell cervical carcinoma; gastric cancer; colon cancer; uterine cervical carcinoma; gall bladder cancer; prostate cancer; breast cancer; testicular germ cell tumors; large cell lymphoma; follicular lymphoma; colorectal cancer; malignant pleural mesothelioma; glioma; thyroid cancer; basal cell carcinoma; T cell lymphoma; t(8;17)-prolyphocytic leukemia; myelodysplastic syndrome; pancreatic cancer; t(5;14)(q35.1;q32.2) leukemia; malignant fibrous histiocytoma; gastrointestinal stromal tumor; and hepatoblastoma. 
     
     
         13 . A method of  claim 11 , wherein the cancer treated is selected from the group consisting of: colorectal; endometrial; ovarian; gastric; and urothelial. 
     
     
         14 . A method of treating cancer in a mismatch repair (MMR) dysfunctional patient in need of such treatment, comprising administering to the patient a pharmaceutically-effective amount of an anti-sense miRNA, wherein the antisense miRNA is antisense to miR-155. 
     
     
         15 . A method of  claim 14 , wherein the cancer treated is selected from the group consisting of: colorectal; endometrial; ovarian; gastric; and urothelial. 
     
     
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         17 . A method of  claim 14 , which further comprises administering a compound selected from the group consisting of: selected from the group consisting of: a chemotherapy drug; a stem cell; AG1478; gefitinib; erlotinib; cetuximab; panitumab; zalutumamab; nimotuzamab; matuzumab; and lapatinib; bevacizumab; cisplatin; carboplatin; procarbazine; mechlorethamine; cyclophosphamide; camptothecin; celecoxib; ifosfamide; melphalan; chlorambucil; busulfan; nitrosurea; dactinomycin; daunorubicin; doxorubicins adriamycin; bleomycin; plicomycin; mitomycin; etoposide; tamoxifen; raloxifene; taxol; taxotere; gemcitabien; navelbine; transplatinum; 5-fluorouracil; vincristin; vinblastin; methotrexate; an EGFR inhibitor; a COX-2 inhibitor; an estrogen receptor binding agent; and a farnesyl-protein transferase inhibitor. 
     
     
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         19 . A method for inducing apoptosis of mismatch repair (MMR) dysfunctional cells, comprising introducing to the MMR dysfunctional cells an apoptosis-effective amount of an anti-sense miRNA, wherein the antisense miRNA is antisense to miR-155. 
     
     
         20 . A method of  claim 19 , wherein cells are dysfunctional due to at least one downregulated core MMR protein selected from the group consisting of: hMSH2; hMSH6; and hMLH1. 
     
     
         21 . A method of  claim 19 , wherein the cells are dysfunctional due to microsatellite instability. 
     
     
         22 . A method of  claim 20 , which further comprises introducing a compound to the MMR dysfunctional cells selected from the group consisting of: a chemotherapy drug; a stem cell; AG1478; gefitinib erlotinib; cetuximab; panitumab; zalutumamab; nimotuzamab; matuzumab; and lapatinib; bevacizumab; cisplatin; carboplatin; procarbazine; mechlorethamine; cyclophosphamide; camptothecin; celecoxib; ifosfamide; melphalan; chlorambucil; busulfan; nitrosurea; dactinomycin; daunorubicin; doxorubicins adriamycin; bleomycin; plicomycin; mitomycin; etoposide; tamoxifen; raloxifene; taxol; taxotere; gemcitabien; navelbine; transplatinum; 5-fluorouracil; vincristin; vinblastin; methotrexate; an EGFR inhibitor; a COX-2 inhibitor; an estrogen receptor binding agent; and a farnesyl-protein transferase inhibitor. 
     
     
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         24 . A method of predicting a clinical outcome of a patient diagnosed with mismatch repair (MMR) dysfunction disease, comprising detecting the expression level of miR-155 in a MMR dysfunction disease cell sample obtained from the patient, wherein a 1.5-fold or greater increase in the level of miR-155 relative to a control, in combination with a MMR dysfunction status predicts the clinical outcome of a decrease in survival of the patient. 
     
     
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         28 . The method of  claim 5 , wherein the MMR dysfunctional cells express at least one dysfunctional core MMR protein selected from the group consisting of: hMSH2, hMSH6 and hMLH1. 
     
     
         29 . A method for restoring a desired expression of at least one core mismatch repair (MMR) protein in a cell in need thereof, comprising administering to the cell an effective amount of at least anti-miR-155 in an sufficient to increase expression of one or more core MMR proteins selected from: human MutS homolog 2 (MSH2), human MutS homolog 6 (MSH6) and human MutL homolog 1 (MLH1). 
     
     
         30 . A method for inducing re-expression of at least one mismatch repair MMR gene in a cell in need thereof, comprising administering to the cell an effective amount of an anti-miR-155 sufficient to induce MMR gene expression selected from one or more of human MutS homolog 2 (MSH2), human MutS homolog 6 (MSH6) and human MutL homolog 1 (MLH1). 
     
     
         31 . A method for inhibiting tumorigenicity both in vitro and in vivo in a subject in need thereof, comprising administering an effective amount of an anti-miR-4155 nucleic acid construct sufficient to target one or more mismatch repair (MMR) gene. 
     
     
         32 . The method of  claim 31 , wherein the subject is a colorectal cancer patient. 
     
     
         33 . A method for increasing expression of a mismatch repair (MMR) protein in a cell in need thereof, the method comprising: transfecting a cell with an anti-mir-155 nucleic acid construct in an amount sufficient to increase expression of one or more MMR proteins selected from: human MutS homolog 2 (MSH2), human MutS homolog 6 (MSH6) and human MutL homolog 1 (MLH1). 
     
     
         34 . A method of epigenetic therapy, comprising administering to a cell in need thereof, a synthetic anti-miR-494, alone or in combination with other treatments, in an amount sufficient to reactivate at least one mismatch repair (MMR) protein expression and normalize aberrant expression patterns of the at least one MMR proteinor TRAIL expression in the cell. 
     
     
         35 . The method of  claim 34 , wherein the cell is a cancer cell. 
     
     
         36 . The method of  claim 35 , wherein the cancer cell is a colorectal cancer cell. 
     
     
         37 . A method of inhibiting tumorigenesis in a subject who has, or is suspected of having, a cancer-related disease in which at least miR-155 is up-regulated in the cancer cells of the subject, relative to control cells, comprising:
 when the miR-155 is up-regulated in the cancer cells, administering to the subject an effective amount of at least anti-miR-155 for inhibiting expression of one or more mismatch repair (MMR) proteins selected from: human MutS homolog 2 (MSH2), human MutS homolog 6 (MSH6) and human MutL homolog 1 (MLH1).the miR-155, such that tumorigenesis is inhibited in the subject.   
     
     
         38 . The method of  claim 37 , wherein the anti-miR-155 comprises an anti-miR-155 nucleic acid construct, or isolated variants or biologically-active fragments or functional equivalents thereof, or antibodies that bind thereto. 
     
     
         39 . A method of up-regulating at least one mismatch repair (MMR) gene in a cell, comprising introducing into the cell an effective amount of a miR-specific inhibitor of at least miR-155 into the cell sufficient to alter expression patterns of one or more proteins encoded by the MMR gene, the mismatch repair (MMR) proteins selected from: human MutS homolog 2 (MSH2), human MutS homolog 6 (MSH6) and human MutL homolog 1 (MLH1). 
     
     
         40 . The method of  claim 39  , wherein the cell is a cancer cell. 
     
     
         41 . The method of  claim 39 , wherein the miR-specific inhibitor is anti-sense miR-155.