Dual targeting of mir-208 and mir-499 in the treatment of cardiac disorders

Abstract

The present invention provides a method of treating or preventing cardiac disorders in a subject in need thereof by inhibiting the expression or function of both miR-499 and miR-208 in the heart cells of the subject. In particular, specific protocols for administering inhibitors of the two miRNAs that achieve efficient, long-term suppression are disclosed. In addition, the invention provides a method for treating or preventing musculoskeletal disorders in a subject in need thereof by increasing the expression or activity of both miR-208 and miR-499 in skeletal muscle cells of the subject.

1 - 25 . (canceled) 
     
     
         26 . A method for treating pathologic cardiac hypertrophy, myocardial infarction, or heart failure in a subject in need thereof comprising administering to the subject a nucleic acid molecule comprising a first miRNA targeting sequence and a second miRNA targeting sequence, wherein the first miRNA targeting sequence is at least partially complementary to a mature miR-208a or miR-208b sequence and the second miRNA targeting sequence is at least partially complementary to a mature miR-499 sequence, and wherein the expression or activity of miR-208a or miR-208b and miR-499 is reduced in the heart cells of the subject following administration of the nucleic acid. 
     
     
         27 . The method of  claim 26 , wherein the nucleic acid is expressed from a vector delivered to the heart cells of the subject. 
     
     
         28 . The method of  claim 26 , wherein the first and second miRNA targeting sequences are separated by one or more spacer nucleotides. 
     
     
         29 . The method of  claim 28 , wherein the first and second miRNA targeting sequences are separated by about 10 to about 50 spacer nucleotides. 
     
     
         30 . The method of  claim 28 , wherein the first and second miRNA targeting sequences are separated by about 5 spacer nucleotides. 
     
     
         31 . The method of  claim 26 , wherein the mature miR-208a sequence is SEQ ID NO: 5. 
     
     
         32 . The method of  claim 31 , wherein the first miRNA targeting sequence is at least 80% complementary to SEQ ID NO: 5. 
     
     
         33 . The method of  claim 31 , wherein the first miRNA targeting sequence is at least 90% complementary to SEQ ID NO: 5. 
     
     
         34 . The method of  claim 31 , wherein the first miRNA targeting sequence is fully complementary to SEQ ID NO: 5. 
     
     
         35 . The method of  claim 26 , wherein the mature miR-208b sequence is SEQ ID NO: 19. 
     
     
         36 . The method of  claim 35 , wherein the first miRNA targeting sequence is at least 80% complementary to SEQ ID NO: 19. 
     
     
         37 . The method of  claim 35 , wherein the first miRNA targeting sequence is at least 90% complementary to SEQ ID NO: 19. 
     
     
         38 . The method of  claim 35 , wherein the first miRNA targeting sequence is fully complementary to SEQ ID NO: 19. 
     
     
         39 . The method of  claim 26 , wherein the mature miR-499 sequence is SEQ ID NO: 14. 
     
     
         40 . The method of  claim 39 , wherein the second miRNA targeting sequence is at least 80% complementary to SEQ ID NO: 14. 
     
     
         41 . The method of  claim 39 , wherein the second miRNA targeting sequence is at least 90% complementary to SEQ ID NO: 14. 
     
     
         42 . The method of  claim 39 , wherein the second miRNA targeting sequence is fully complementary to SEQ ID NO: 14. 
     
     
         43 . The method of  claim 26 , wherein the nucleic acid comprises at least one sugar and/or backbone modification. 
     
     
         44 . The method of  claim 43 , wherein said at least one sugar modification is a bicyclic sugar nucleoside modification, 2′-O-alkyl modification, or 2′-fluoro modification. 
     
     
         45 . The method of  claim 44 , wherein the bicyclic sugar nucleoside modification is a locked nucleic acid. 
     
     
         46 . The method of  claim 43 , wherein said at least one backbone modification is a phosphorothioate linkage. 
     
     
         47 . The method of  claim 26 , wherein the first miRNA targeting sequence and/or the second miRNA targeting sequence are about 8 to about 18 nucleotides in length. 
     
     
         48 . The method of  claim 26 , wherein the first miRNA targeting sequence and/or the second miRNA targeting sequence are about 12 to about 16 nucleotides in length.