STAT5a AND ITS FUNCTIONAL TUMOR SUPPRESSOR ANALOGS FOR TREATMENT OF MALIGNANCIES EXPRESSING NPM/ALK AND OTHER ONCOGENIC KINASES

Abstract

The invention provides methods of inhibiting epigenetic gene silencing in a cell expressing NPM/ALK or decreasing NPM/ALK content in a cell, by contacting a cell with an agent capable of increasing the concentration of Stat5a protein or its functional analog. Further, the invention provides a method of treating malignancies expressing oncogenic kinase by administering to a patient affected with a malignancy an agent capable of increasing the concentration of Stat5a protein or its epigenetically silenced functional tumor suppressor analog in a malignant cell. Finally, it provides a method to diagnose malignancy and monitor patient's response to therapy by analysis of the degree of DNA methylation of the gene encoding for Stat5a or its analog, their mRNA, or protein.

1 .- 10 . (canceled) 
     
     
         11 . A method of decreasing NPM/ALK, or another oncogenic -kinase content in a cell, comprising the step of contacting a cell expressing NPM/ALK or another oncogenic -kinase, with an effective amount of an agent capable of increasing the amount of Stat5a or its functional analog, thereby down-regulating the kinase gene expression in the cell. 
     
     
         12 . The method of  claim 11 , whereby the agent is Stat5a protein, a Stat5a mRNA, Stat5a DNA or a combination thereof. 
     
     
         13 . The method of  claim 12 , further comprising a siRNA, a polyamide, a triple-helix-forming agent, an antisense RNA, a synthetic peptide nucleic acids (PNAs), an agRNA, a LNA/DNA copolymer, a small molecule chemical compound, or a combination thereof, specific against a NPM/ALK, another form of ALK, another chimeric tyrosine kinase, or another oncogenic kinase. 
     
     
         14 . A method of treating malignancies expressing an ALK +  chimeric tyrosine kinase in a subject, comprising the step of administering to the subject a composition comprising a therapeutically effective amount of a small molecule chemical compound, wherein the small molecule chemical compound is a DNA Methyltransferase (DNMT) inhibitor. 
     
     
         15 . The method of  claim 14 , whereby the agent is capable of increasing Stat5a expression. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 14 , whereby the composition further comprises an agent capable of inhibiting the expression or function of a methylated DNA capping family (MBD) protein, Stat3 or another transcriptional repressor mediating epigenetic silencing of STAT5a or its functional analog, or their combination. 
     
     
         18 . The method of  claim 17 , wherein the methylated DNA capping family (MBD) protein is Methylation binding domain protein. 
     
     
         19 . The method of  claim 17 , whereby the agent capable of inhibiting the function of a methylated DNA capping family (MBD) protein is an antibody or a fragment thereof, specific against a methylated DNA capping family (MBD) protein. 
     
     
         20 . The method of  claim 19 , whereby the agent capable of inhibiting the function of Methylation binding domain family protein, or Stat3 or another transcriptional repressor mediating epigenetic silencing of STAT5a or its functional analog, is a small molecule chemical compound. 
     
     
         21 . The method of  claim 19 , whereby the agent capable of inhibiting the expression of a methylated DNA capping family (MBD) protein is a siRNA, a polyamide, a triple-helix-forming agent, an antisense RNA, a synthetic peptide nucleic acids (PNAs), an agRNA, a LNA/DNA copolymer, a small molecule chemical compound, or a combination thereof, specific against a nucleotide sequence encoding a methylated DNA capping family (MBD) protein, or Stat3. 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 14 , whereby the DNMT inhibitor is 5′-aza-2′-deoxycytidine, or 5-azacytidine. 
     
     
         24 . The method of  claim 14 , whereby the malignancy expressing chimeric tyrosine kinase is lymphoma, leukemia, sarcoma, or carcinoma. 
     
     
         25 .- 33 . (canceled) 
     
     
         34 . A method of preventing, inhibiting or suppressing epigenetic silencing of transcriptional expression of a tumor suppressor in a subject, comprising the step of contacting the subject with a therapeutically effective amount of a composition comprising an agent capable of inhibiting the expression or function of a transcription inhibitor, a Methylation binding domain protein, a DNMT or their combination. 
     
     
         35 . The method of  claim 34 , whereby the transcription factor is Stat3. 
     
     
         36 . The method of  claim 34 , whereby the Methylation binding domain protein is a MeCP or related protein. 
     
     
         37 . The method of  claim 34 , whereby the agent is a siRNA, a polyamide, a triple-helix-forming agent, an antisense RNA, a synthetic peptide nucleic acids (PNAs), an agRNA, a LNA/DNA copolymer, a small molecule chemical compound, or a combination thereof, specific against a nucleotide sequence encoding a transcription factor, a Methylation binding domain protein, a DNMT or their combination. 
     
     
         38 . The method of  claim 34 , whereby the DNMT inhibitor is 5′-aza-2′-deoxycytidine, or 5-azacytidine. 
     
     
         39 . The method of  claim 34 , whereby the tumor suppressor gene is Stat5a, SHP-1 phosphatase, or their combination. 
     
     
         40 . A method of treating, inhibiting, suppressing, or ameliorating symptoms associated with a T-cell lymphoma in a subject, comprising the step of administering to the subject a therapeutically effective amount of a composition comprising an agent capable of increasing the expression or function of Stat5a protein or its analog thereby treating, inhibiting, suppressing, or ameliorating symptoms associated with the T-cell lymphoma in the subject. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . The method of  claim 41 , whereby the agent is capable of increasing Stat5a expression. 
     
     
         44 . The method of  claim 51 , whereby the agent is Stat5a mRNA. 
     
     
         45 . The method of  claim 41 , whereby the composition further comprises an agent capable of inhibiting the expression or function of a transcription factor, a DNMT, a methylated DNA capping family (MBD) protein or their combination. 
     
     
         46 . The method of  claim 45 , whereby the methylated DNA capping family (MBD) protein is Methylation binding domain. 
     
     
         47 . The method of  claim 45 , whereby the agent capable of inhibiting the function of a transcription factor, a Methylation binding domain protein, a DNMT or their combination, is an antibody or a fragment thereof, specific against the transcription factor, Methylation binding domain protein, DNMT or their combination. 
     
     
         48 . The method of  claim 45 , whereby the agent capable of inhibiting the function of a transcription factor, a Methylation binding domain protein, a DNMT or their combination is a small molecule chemical compound. 
     
     
         49 . The method of  claim 45 , whereby the agent capable of inhibiting the expression of a transcription factor, a Methylation binding domain protein, a DNMT or their combination, is a siRNA, a polyamide, a triple-helix-forming agent, an antisense RNA, a synthetic peptide nucleic acids (PNAs), an agRNA, a LNA/DNA copolymer, a small molecule chemical compound, or a combination thereof, specific against a nucleotide sequence encoding a methylated DNA capping family (MBD) protein. 
     
     
         50 . The method of  claim 41 , whereby the agent is a DNMT inhibitor. 
     
     
         51 . The method of claim  57 , whereby the DNMT inhibitor is 5′-aza-2′-deoxy-cytidine (5′-aza). 
     
     
         52 . The method of  claim 41 , whereby administering is via contacting ex-vivo, a cell of the subject with the composition and reinserting the cell to the subject. 
     
     
         53 . The method of  claim 41 , whereby the transcription factor is Stat3. 
     
     
         54 . The method of  claim 41 , whereby the T-cell lymphoma is an anaplastic large cell lymphoma. 
     
     
         55 .- 79 . (canceled)