Recombinant adeno-associated virus virions for the treatment of lysosomal disorders

Abstract

AAV expression vectors and recombinant virions produced using these vectors, which include genes coding for enzymes defective or missing in lysosomal storage disorders, are described. These recombinant AAV virions are useful in the treatment of a variety of lysosomal storage disorders and the methods described herein provide for long-term, sustained expression of the defective or missing enzyme.

1 . A method of delivering a recombinant adeno-associated virus (AAV) virion to a mammalian subject to treat a lysosomal storage disease, said method comprising: 
 (a) providing a recombinant AAV virion which comprises a nucleic acid molecule, said nucleic acid molecule comprising a gene encoding a protein defective or missing in the lysosomal storage disease operably linked to control elements capable of directing the in vivo transcription and translation of said gene; and    (b) delivering said recombinant AAV virion to the bloodstream of said subject, whereby said gene is expressed at a level which provides a therapeutic effect in said mammalian subject.    
     
     
         2 . The method of  claim 1 , wherein the recombinant AAV virion is delivered intravenously.  
     
     
         3 . The method of  claim 2 , wherein the recombinant AAV virion is delivered to the portal vein.  
     
     
         4 . The method of  claim 1 , wherein the recombinant AAV virion is delivered intraarterially.  
     
     
         5 . The method of  claim 4 , wherein the recombinant AAV virion is delivered to the hepatic artery.  
     
     
         6 . The method of  claim 1 , wherein the gene is expressed in the liver of said subject.  
     
     
         7 . The method of  claim 1 , wherein the lysosomal storage disease is MPS VII and the gene encodes β-glucuronidase.  
     
     
         8 . A method of delivering a gene encoding a protein defective or missing in a lysosomal storage disease to a mammalian subject, said method comprising: 
 (a) providing a recombinant adeno-associated virus (AAV) virion which comprises a nucleic acid molecule, said nucleic acid molecule comprising a gene encoding a protein defective or missing in the lysosomal storage disease operably linked to control elements capable of directing the in vivo transcription and translation of said gene; and    (b) delivering said recombinant AAV virion to the liver or brain of said subject, whereby said gene is expressed in the liver or brain at a level which provides a therapeutic effect in said mammalian subject.    
     
     
         9 . The method of  claim 8 , wherein the recombinant AAV virion is delivered to the liver of said subject.  
     
     
         10 . The method of  claim 8 , wherein the recombinant AAV virion is delivered to the brain of said subject.  
     
     
         11 . The method of  claim 10 , wherein the recombinant AAV virion is delivered to the brain by intrathecal injection.  
     
     
         12 . The method of  claim 8 , wherein the lysosomal storage disease is MPS VII and the gene encodes β-glucuronidase.  
     
     
         13 . A method of delivering a recombinant adeno-associated virus (AAV) virion to a mammalian subject to treat MPS VII, said method comprising: 
 (a) providing a recombinant AAV virion which comprises a nucleic acid molecule, said nucleic acid molecule comprising a gene encoding β-glucuronidase operably linked to control elements capable of directing the in vivo transcription and translation of said gene; and    (b) delivering said recombinant AAV virion to said subject by intrathecal injection, whereby said gene is expressed at a level which provides a therapeutic effect in said mammalian subject.    
     
     
         14 . A recombinant adeno-associated virus vector comprising a gene encoding β-glucuronidase operably linked to control elements capable of directing the in vivo transcription and translation of said gene in a mammalian cell.  
     
     
         15 . A recombinant adeno-associated virus (AAV) virion comprising a nucleic acid molecule, said nucleic acid molecule comprising a gene encoding β-glucuronidase operably linked to control elements capable of directing the in vivo transcription and translation of said gene in a mammalian cell.  
     
     
         16 . A method of producing a recombinant adeno-associated virus (AAV) virion comprising: 
 (a) transfecting a host cell with (i) an AAV vector comprising a gene encoding β-glucuronidase operably linked to control elements capable of directing the in vivo transcription and translation of said gene in a mammalian cell; (ii) AAV helper functions; and (iii) AAV accessory functions,    wherein said transfecting is done under conditions that allow the formation of a recombinant AAV virion; and    (b) purifying the recombinant AAV virion from the host cell.