Adeno-associated virus virions with variant capsid and methods of use thereof

Abstract

The present disclosure provides adeno-associated virus (AAV) virions with altered capsid protein, where the AAV virions exhibit greater infectivity of retinal cells compared to wild-type AAV. The present disclosure further provides methods of delivering a gene product to a retinal cell in an individual, and methods of treating ocular disease.

1 .- 30 . (canceled) 
     
     
         31 . A recombinant adeno-associated virus (rAAV) virion comprising:
 a) a variant AAV capsid protein, wherein the variant AAV capsid protein comprises an amino acid substitution at amino acid 451 of the AAV6 capsid sequence as set forth in SEQ ID NO:1, or the corresponding position in another AAV parental serotype, wherein the AAV capsid protein does not comprise an amino acid sequence present in a naturally occurring AAV capsid protein; and   b) a heterologous nucleic acid comprising a nucleotide sequence encoding a heterologous gene product.   
     
     
         32 . The rAAV virion of  claim 31 , wherein heterologous gene product is a nucleic acid gene product. 
     
     
         33 . The rAAV virion of  claim 32 , wherein the nucleic acid gene product is an interfering RNA, a ribozyme, an antisense nucleic acid, or an aptamer. 
     
     
         34 . The rAAV virion of  claim 33 , wherein the interfering RNA or the aptamer reduces the level of an angiogenic factor in the retinal cell. 
     
     
         35 . The rAAV virion of  claim 31 , wherein the heterologous gene product is a polypeptide. 
     
     
         36 . The rAAV virion of  claim 35 , wherein the polypeptide is a neuroprotective polypeptide. 
     
     
         37 . The rAAV virion of  claim 35 , wherein the polypeptide is glial derived neurotrophic factor, fibroblast growth factor 2, nurturin, ciliary neurotrophic factor, nerve growth factor, brain derived neurotrophic factor, epidermal growth factor, a soluble vascular endothelial growth factor (VEGF) receptor, an anti-VEGF antibody, or Sonic hedgehog. 
     
     
         38 . The rAAV virion of  claim 35 , wherein the polypeptide is an anti-angiogenic polypeptide. 
     
     
         39 . The rAAV virion of  claim 31 , wherein the parental AAV capsid protein is wild-type AAV6 capsid protein. 
     
     
         40 . The rAAV virion of  claim 31 , wherein the variant AAV capsid protein comprises an amino acid change at AAV6 position 532 or the corresponding position in another AAV parental serotype. 
     
     
         41 . The rAAV virion of  claim 31 , wherein the amino acid substitution at amino acid 451 of AAV6, or the corresponding position in another AAV parental serotype, is an aspartic acid. 
     
     
         42 . The rAAV virion of  claim 31 , wherein the variant capsid protein comprises from 1 to 10 amino acid differences compared to a wild-type AAV capsid protein. 
     
     
         43 . A pharmaceutical composition comprising:
 a) a recombinant adeno-associated virus (rAAV) virion according to  claim 31 ; and   b) a pharmaceutically acceptable carrier, diluent, excipient, or buffer.   
     
     
         44 . A method of delivering a gene product to a retinal cell in an individual, the method comprising administering to the individual a recombinant adeno-associated virus (rAAV) virion according to  claim 31 . 
     
     
         45 . The method of  claim 44 , wherein the gene product is a polypeptide. 
     
     
         46 . The method of  claim 45 , wherein the polypeptide is a neuroprotective factor or an anti-angiogenic factor. 
     
     
         47 . The method of  claim 45 , wherein the polypeptide is glial derived neurotrophic factor, fibroblast growth factor 2, nurturin, ciliary neurotrophic factor, nerve growth factor, brain derived neurotrophic factor, epidermal growth factor, a soluble vascular endothelial growth factor (VEGF) receptor, an anti-VEGF antibody, or Sonic hedgehog. 
     
     
         48 . The method of  claim 44 , wherein the gene product is a nucleic acid. 
     
     
         49 . A method of treating a retinal disease, the method comprising administering to an individual in need thereof an effective amount of a recombinant adeno-associated virus (rAAV) virion according to  claim 31 . 
     
     
         50 . The method of  claim 49 , wherein said administering is by intraocular injection. 
     
     
         51 . The method of  claim 49 , wherein said administering is by intravitreal injection.