Methods and Compositions for use in Gene Therapy for Treatment of Hemophilia

Abstract

Provided are compositions of viruses, the viruses including a recombinant adeno-associated virus vector comprising at least two adeno-associated virus inverted terminal repeats, a promoter/regulatory sequence, isolated DNA encoding a blood coagulation factor and accompanying 5′ and 3′ untranslated regions and a transcription termination sequence.

1 . A composition comprising a virus, said virus comprising a recombinant adeno-associated virus vector comprising at least two adeno-associated virus inverted terminal repeats, a promoter/regulatory sequence, isolated DNA encoding a blood coagulation factor and accompanying 5′ and 3′ untranslated regions and a transcription termination sequence. 
     
     
         2 . The composition of  claim 1 , wherein said blood coagulation factor comprises Factor VII, Factor VIII, Factor X, Factor XI, Factor XIII, or Protein C. 
     
     
         3 . A method of treating a blood coagulation factor deficiency in a mammal comprising:
 (a) providing a recombinant adeno-associated virus vector, said vector comprising a nucleic acid encoding a blood coagulation factor operably linked to an expression control element; and   (b) administering an amount of said virus vector to a mammal wherein said blood coagulation factor is expressed at levels having a therapeutic effect on said mammal and wherein said therapeutic effect is an increase in coagulation of blood.   
     
     
         4 . The method of  claim 3 , wherein said blood coagulation factor is Factor VII, Factor VIII, Factor X, Factor XI, Factor XIII, or Protein C. 
     
     
         5 . A method for expressing Factor IX in a mammal comprising injecting a recombinant adeno-associated virus vector, said vector comprising a nucleic acid encoding Factor IX operably linked to an expression control element, into the muscle tissue of said mammal. 
     
     
         6 . The method of  claim 5  wherein said vector is injected at a single site per dose. 
     
     
         7 . The method of  claim 5  wherein said vector is injected at multiple sites. 
     
     
         8 . The method of  claim 5  wherein said vector further comprises at least two adeno-associated virus inverted terminal repeats, a promoter/regulatory sequence, isolated DNA encoding Factor IX and accompanying 5′ and 3′ untranslated regions and a transcription termination sequence. 
     
     
         9 . The method of  claim 5  wherein said vector further comprises a portion of intron I of the Factor IX gene. 
     
     
         10 . The method of  claim 9  wherein said portion of intron I of said Factor IX gene is from about 0.3 kb to about 1.7 kb. 
     
     
         11 . The method of  claim 5  wherein said Factor IX is human Factor IX. 
     
     
         12 . The method of  claim 5  wherein nucleic acid encoding Factor IX comprises a mutation encoding an alanine residue in place of lysine in the fifth amino acid position from the beginning of mature Factor IX, wherein said mutation reduces the ability of said Factor IX to bind to collagen IV.