Modulating phosphatase activity in cardiac cells

Abstract

Expression of a phosphatase inhibitor in heart cells can be used to treat cardiac disorders, e.g., heart failure. Decreasing phosphatase activity can improve β-adrenergic responsiveness.

1 . An adeno-associated virus (AAV) vector comprising a nucleic acid sequence encoding a polypeptide consisting of amino acids 1-65 of SEQ ID NO: 2, wherein threonine at amino acid 35 of SEQ ID NO: 2 is replaced with an aspartic acid, and wherein said nucleic acid sequence is operably linked to a promoter capable of directing expression in the heart. 
     
     
         2 . The AAV vector of  claim 1 , wherein the promoter is a tissue specific promoter, a smooth muscle specific promoter, a cardiac specific promoter, or a viral promoter. 
     
     
         3 . The AAV vector of  claim 2 , wherein the tissue specific promoter is a cardiac troponin T promoter, myosin heavy chain or the myosin light chain promoter, cardiac myosin promoter, troponin T promoter, BNP promoter, alpha actin promoter, or a SM22a promoter. 
     
     
         4 . The AAV vector of  claim 1 , wherein the promoter is a constitutive promoter. 
     
     
         5 . The AAV vector of  claim 1 , wherein the promoter is expressed in a subset of tissues, at least one of which is a cardiac muscle. 
     
     
         6 . The AAV vector of  claim 1 , wherein the promoter is a CMV promoter. 
     
     
         7 . The AAV vector of  claim 1 , wherein the AAV is selected from the group consisting of adeno-associated virus-1 (AAV1), AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 and AAV9. 
     
     
         8 . A pharmaceutical composition comprising:
 a) an adeno-associated virus (AAV) vector comprising a nucleic acid sequence encoding a polypeptide consisting of amino acids 1-65 of SEQ ID NO: 2, wherein threonine at amino acid 35 of SEQ ID NO: 2 is replaced with an aspartic acid, and wherein said nucleic acid sequence is operably linked to a promoter capable of directing expression in the heart; and   b) a pharmaceutically acceptable carrier.   
     
     
         9 . The pharmaceutical composition of  claim 8 , wherein the promoter is a tissue specific promoter, a smooth muscle specific promoter, a cardiac specific promoter, or a viral promoter. 
     
     
         10 . The pharmaceutical composition of  claim 9 , wherein the tissue specific promoter is a cardiac troponin T promoter, myosin heavy chain or the myosin light chain promoter, cardiac myosin promoter, troponin T promoter, BNP promoter, alpha actin promoter, or a SM22a promoter. 
     
     
         11 . The pharmaceutical composition of  claim 8 , wherein the promoter is a constitutive promoter. 
     
     
         12 . The pharmaceutical composition of  claim 8 , wherein the promoter is expressed in a subset of tissues, at least one of which is a cardiac muscle. 
     
     
         13 . The pharmaceutical composition of  claim 8 , wherein the promoter is a CMV promoter. 
     
     
         14 . The pharmaceutical composition of  claim 8 , wherein the AAV is selected from the group consisting of adeno-associated virus-1 (AAV1), AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 and AAV9. 
     
     
         15 . A container comprising an adeno-associated virus (AAV) vector comprising a nucleic acid sequence encoding a polypeptide consisting of amino acids 1-65 of SEQ ID NO: 2, wherein threonine at amino acid 35 of SEQ ID NO: 2 is replaced with an aspartic acid, and wherein said nucleic acid sequence is operably linked to a promoter capable of directing expression in the heart. 
     
     
         16 . The container of  claim 15 , further comprising a pharmaceutically acceptable carrier. 
     
     
         17 . The container of  claim 16 , wherein the promoter is a tissue specific promoter, a smooth muscle specific promoter, a cardiac specific promoter, or a viral promoter. 
     
     
         18 . The container of  claim 16 , wherein the tissue specific promoter is a cardiac troponin T promoter, myosin heavy chain or the myosin light chain promoter, cardiac myosin promoter, troponin T promoter, BNP promoter, alpha actin promoter, or a SM22a promoter. 
     
     
         19 . The container of  claim 16 , wherein the promoter is a constitutive promoter. 
     
     
         20 . The container of  claim 16 , wherein the promoter is expressed in a subset of tissues, at least one of which is a cardiac muscle. 
     
     
         21 . The container of  claim 16 , wherein the promoter is a CMV promoter. 
     
     
         22 . The container of  claim 16 , wherein the AAV is selected from the group consisting of adeno-associated virus-1 (AAV1), AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 and AAV9. 
     
     
         23 . The container of  claim 16 , comprising at least 1×10 9  viral genomes. 
     
     
         24 . The container of  claim 16 , comprising at least 1×10 10  viral genomes. 
     
     
         25 . The container of  claim 16 , comprising at least 1×10 13  viral genomes. 
     
     
         26 . The container of  claim 16 , comprising at least 1×10 14  viral genomes. 
     
     
         27 . The container of  claim 16 , comprising at least 1×10 15  viral genomes. 
     
     
         28 . The container of  claim 16 , comprising at least 1×10 16  viral genomes. 
     
     
         29 . The container of  claim 16 , comprising between 1×10 9  and 1×10 18 , or between 1×10 11  and 1×10 16  viral genomes. 
     
     
         30 . The container of  claim 16  that is a syringe.