Methods for delivering DNA to muscle cells using recombinant adeno-associated virus virions vectors

Abstract

The use of recombinant adeno-associated virus (AAV) virions for delivery of DNA molecules to muscle cells and tissue is disclosed. The invention allows for the direct, in vivo injection of recombinant AAV virions into muscle tissue, e.g., by intramuscular injection, as well as for the in vitro transduction of muscle cells which can subsequently be introduced into a subject for treatment. The invention provides for sustained, high-level expression of the delivered gene and for in vivo secretion of the therapeutic protein from transduced muscle cells such that systemic delivery is achieved.

1 . A method of delivering a selected gene to a muscle cell or tissue, said method comprising: 
 (a) providing a recombinant adeno-associated virus (AAV) virion which comprises an AAV vector, said AAV vector comprising said selected gene operably linked to control elements capable of directing the in vivo transcription and translation of said selected gene; and    (b) introducing said recombinant AAV virion into said muscle cell or tissue.    
     
     
         2 . The method of  claim 1 , wherein said muscle cell or tissue is derived from skeletal muscle.  
     
     
         3 . The method of  claim 1 , wherein said muscle cell or tissue is derived from smooth muscle.  
     
     
         4 . The method of  claim 1 , wherein said muscle cell or tissue is derived from cardiac muscle.  
     
     
         5 . The method of  claim 1 , wherein said muscle cell is a skeletal myoblast.  
     
     
         6 . The method of  claim 1 , wherein said muscle cell is a skeletal myocyte.  
     
     
         7 . The method of  claim 1 , wherein said muscle dell is a cardiomyocyte.  
     
     
         8 . The method of  claim 1 , wherein said recombinant AAV virion is introduced into said muscle cell in vivo.  
     
     
         9 . The method of  claim 8 , wherein said recombinant AAV virion is introduced by intramuscular injection.  
     
     
         10 . The method of  claim 1 , wherein said recombinant AAV virion is introduced into said muscle cell in vitro.  
     
     
         11 . The method of  claim 1 , wherein said selected gene encodes a therapeutic protein.  
     
     
         12 . The method of  claim 11 , wherein said protein is acid α-glucosidase.  
     
     
         13 . A muscle cell or tissue transduced with a recombinant AAV virion which comprises an AAV vector, said AAV vector comprising a selected gene operably linked to control elements capable of directing the in vivo transcription and translation of said selected gene.  
     
     
         14 . The muscle cell of  claim 13 , wherein said cell is a skeletal myoblast.  
     
     
         15 . The muscle cell of  claim 13 , wherein said cell is a skeletal myocyte.  
     
     
         16 . The muscle cell of  claim 13 , wherein said cell is a cardiomyocyte.  
     
     
         17 . The muscle cell of  claim 13 , wherein said selected gene encodes a therapeutic protein.  
     
     
         18 . The muscle cell of  claim 17 , wherein said selected gene encodes acid α-glucosidase.  
     
     
         19 . A method of treating an acquired or inherited disease in a mammalian subject comprising introducing into a muscle cell or tissue of said subject a therapeutically effective amount of a pharmaceutical composition which comprises (a) a pharmaceutically acceptable excipient; and (b) recombinant AAV virions, wherein said recombinant AAV virions comprise an AAV vector, said AAV vector comprising a selected gene operably linked to control elements capable of directing the transcription and translation of said selected gene when present in said subject, wherein said introducing is done in vivo.  
     
     
         20 . A method of treating an acquired or inherited disease in a mammalian subject comprising: 
 (a) introducing a recombinant AAV virion into a muscle cell or tissue in vitro to produce a transduced muscle cell, wherein said recombinant AAV virion comprises an AAV vector, said AAV vector comprising a selected gene operably linked to control elements capable of directing the transcription and translation of said selected gene when present in said subject; and    (b) administering to said subject a therapeutically effective amount of a composition comprising a pharmaceutically acceptable excipient and the transduced muscle cells from step (a).    
     
     
         21 . A method for delivering a therapeutically effective amount of a protein systemically to a mammalian subject comprising introducing into a muscle dell or tissue of said subject a pharmaceutical composition which comprises (a) a pharmaceutically acceptable excipient; and (b) recombinant AAV virions, wherein said recombinant AAV virions comprise an AAV vector, said AAV vector comprising a selected gene operably linked to control elements capable of directing the transcription and translation of said selected gene when present in said subject, wherein said introducing is done in vivo.  
     
     
         22 . A method for delivering a therapeutically effective amount of a protein systemically to a mammalian subject comprising: 
 (a) introducing a recombinant AAV virion into a muscle cell or tissue in vitro to produce a transduced muscle cell, wherein said recombinant AAV virion comprises an AAV vector, said AAV vector comprising a selected gene operably linked to control elements capable of directing the transcription and translation of said selected gene when present in said subject; and    (b) administering to said subject a therapeutically effective amount of a composition comprising a pharmaceutically acceptable excipient and the transduced muscle cells from step (a).    
     
     
         23 . An adeno-associated virus (AAV) vector comprising a gene encoding acid α-glucosidase operably linked to control elements capable of directing the in vivo transcription and translation of said gene.  
     
     
         24 . A recombinant adeno-associated virus (AAV) virion which comprises the AAV vector of  claim 23.