Recombinant adeno-associated vector-mediated delivery of B-domain-deleted factor VIII constructs for the treatment of hemophilia

Abstract

One form of a composition has two types of recombinant adeno-associated virus. The first type encodes a portion of Factor VIII operably linked to an expression control element; and the second type encodes a different portion of Factor VIII operably linked to an expression control element. The first and second nucleotide sequences collectively encode a functional Factor VIII protein. Another form of the composition is a recombinant adeno-associated virus containing a nucleotide sequence encoding functional Factor VIII light or heavy chain operably linked to a tissue-specific promoter.

What is claimed is:  
     
         1 . A method of treating hemophilia in a mammal, comprising: 
 providing a pharmaceutical composition comprising recombinant adeno-associated virus virions, said virions comprising a nucleotide sequence encoding a Factor VIII protein lacking at least a portion of the B domain, said nucleotide sequence operably linked to expression control elements; and    administering said pharmaceutical composition to a mammal under conditions that result in the expression of the Factor VIII protein at a level that provides a therapeutic effect in said mammal.    
     
     
         2 . The method of  claim 9 , wherein said Factor VIII protein is expressed in the liver.  
     
     
         3 . The method of  claim 9 , wherein said recombinant adeno-associated virus virions are administered to the liver.  
     
     
         4 . The method of  claim 9 , wherein said expression control elements comprise a tissue-specific promoter.  
     
     
         5 . The method of  claim 12  wherein said expression control elements comprise a liver-specific promoter.  
     
     
         6 . The method of  claim 9  wherein said expression control elements comprise a human growth hormone polyadenylation sequence.  
     
     
         7 . The method of  claim 9 , wherein said recombinant adeno-associated virus virions are administered via intravenous administration.  
     
     
         8 . The method of claim  15 , wherein said intravenous administration is via the portal vein.  
     
     
         9 . The method of  claim 9 , wherein said recombinant adeno-associated virus virions are administered via intraarterial administration.  
     
     
         10 . The method of claim  17 , wherein said recombinant adeno-associated virus virions are administered via the hepatic artery.  
     
     
         11 . The method of  claim 9 , wherein said nucleotide sequence encoding Factor VIII comprises a light chain and a heavy chain and wherein said light chain and heavy chain are operably linked by a junction.  
     
     
         12 . The method of claim  19 , wherein said nucleotide sequence is SEQ ID 13, such that said junction has the amino acid sequence Ser-Phe.  
     
     
         13 . The method of claim  19 , wherein said nucleotide sequence is SEQ ID 14, such that said junction has the amino acid sequence Ser-Phe-Ser-Gln-Asn-Pro-Pro-Val-Leu-Lys-Arg-His-Gln-Arg.  
     
     
         14 . The method of claim  19 , wherein said expression control elements comprise a liver-specific promoter, and wherein said recombinant adeno-associated virus virions are administered to the liver of said mammal.