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, when administered via intravitreal injection, 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 .- 23 . (canceled) 
     
     
         24 . A variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises an insertion of from about 5 amino acids to about 11 amino acids wherein the amino acid insertion is in the GH loop of a native AAV capsid, wherein the insertion is a peptide of Formula I, Formula II, Formula III, or Formula IV. 
     
     
         25 .- 27 . (canceled) 
     
     
         28 . The variant adeno-associated virus (AAV) capsid protein of  claim 24 , wherein the variant AAV capsid protein comprises an insertion of from about 5 amino acids to about 11 amino acids wherein the amino acid insertion is in the GH loop of a native AAV capsid, wherein the insertion is a peptide of Formula IV. 
     
     
         29 . The variant adeno-associated virus capsid protein of  claim 28 , wherein the amino acid insertion comprises the amino acid sequence Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 , wherein:
 each of Y 1 -Y 4 , if present, is independently selected from Ala, Leu, Gly, Ser, and Thr;   X 1 , if present, is an uncharged amino acid;   X 2  is an uncharged amino acid;   X 3  is a negatively charged amino acid; or is selected from Glu, Thr, Gly, Asp, or Pro;   X 4  is selected from Thr, Ile, Gly, Lys, Asp, and Gln;   X 5  is a polar amino acid, an alcohol (an amino acid having a free hydroxyl group); or is selected from Thr, Ser, Val, and Ala;   X 6  is a positively charged amino acid; or is selected from Arg, Val, Lys, Pro, Thr, and Asn; and   X 7 , if present, is a positively charged amino acid or selected from Pro, Gly, Phe, Asn and Arg.   
     
     
         30 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 3  is a negatively charged amino acid. 
     
     
         31 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 3  is selected from Glu, Thr, Gly, Asp, or Pro. 
     
     
         32 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 3  is Asp. 
     
     
         33 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 4  is Gln. 
     
     
         34 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 5  is a polar amino acid. 
     
     
         35 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 5  is an alcohol (an amino acid having a free hydroxyl group). 
     
     
         36 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 5  is a hydrophobic amino acid. 
     
     
         37 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 5  is selected from Thr, Ser, Val, and Ala. 
     
     
         38 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 5  is Thr. 
     
     
         39 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 6  is a positively charged amino acid. 
     
     
         40 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 6  is selected from Arg, Val, Lys, Pro, Thr, and Asn. 
     
     
         41 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 6  is Lys. 
     
     
         42 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 7 , if present, is a positively charged amino acid. 
     
     
         43 . The variant adeno-associated virus capsid protein of  claim 29 , wherein X 7 , if present, is selected from Pro, Gly, Phe, Asn and Arg. 
     
     
         44 . The variant adeno-associated virus capsid protein of  claim 29 , wherein the amino acid insertion is AVDTTKR (SEQ ID NO: 59). 
     
     
         45 . The variant adeno-associated virus capsid protein of  claim 29 , wherein the amino acid insertion is inserted following any of amino acids in positions 570-671 in VP1 of AAV2 or a corresponding position in another wild-type AAV serotype or AAV variant. 
     
     
         46 . The variant adeno-associated virus capsid protein of  claim 29 , wherein the amino acid insertion is inserted between amino acids 587 and 588 relative to SEQ ID NO: 1. 
     
     
         47 . A variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises an amino acid insertion selected from NETITRP (SEQ ID NO: 14), KAGQANN (SEQ ID NO: 15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO: 57), RAGGSVG (SEQ ID NO: 58), AVDTTKR (SEQ ID NO: 59), or STGKVPN (SEQ ID NO: 60), wherein the amino acid insertion is in the GH loop of a native AAV capsid. 
     
     
         48 . The variant adeno-associated virus capsid protein of  claim 47 , wherein the amino acid insertion is inserted following any of amino acids in positions 570-671 in VP1 of AAV2 or a corresponding position in another wild-type AAV serotype or AAV variant. 
     
     
         49 . The variant adeno-associated virus capsid protein of  claim 47 , wherein the amino acid insertion is inserted between amino acids 587 and 588 relative to SEQ ID NO: 1.