Adeno-associated virus virions with variant capsid and methods of use thereof
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. A recombinant adeno-associated virus (rAAV) virion comprising:
a) a variant AAV capsid protein, wherein the variant AAV capsid protein comprises an insertion of a heterologous peptide in the capsid protein GH loop relative to a corresponding parental AAV capsid protein, and wherein the variant capsid protein confers increased infectivity of a retinal cell compared to the infectivity of the retinal cell by a control AAV virion comprising the corresponding parental AAV capsid protein; and
b) a heterologous nucleic acid comprising a nucleotide sequence encoding a heterologous gene product,
wherein the heterologous peptide comprises the amino acid sequence:
LAPDSTTRSA (SEQ ID NO:53).
2. The rAAV virion of claim 1 , wherein the insertion site is between amino acids corresponding to amino acids 570 and 611 of VP1 of AAV2, or the corresponding position in the capsid protein of another AAV serotype.
3. The rAAV virion of claim 1 , wherein the insertion site is located between amino acids corresponding to amino acids 587 and 588 of VP1 of AAV2, or the corresponding position in the capsid protein of another AAV serotype.
4. The rAAV virion of claim 1 , wherein gene product is an interfering RNA or an aptamer.
5. The rAAV virion of claim 1 , wherein the gene product is a polypeptide.
6. The rAAV virion of claim 5 , wherein the polypeptide is a neuroprotective polypeptide, an anti-angiogenic polypeptide, a polypeptide that enhances function of a retinal cell, or an RNA-guided endonuclease.
7. A pharmaceutical composition comprising:
a) a recombinant adeno-associated virus virion of claim 1 ; and
b) a pharmaceutically acceptable excipient.
8. 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 claim 1 .
9. A method of treating an ocular 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 1 .
10. An isolated nucleic acid comprising a nucleotide sequence that encodes a variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises an insertion of a heterologous peptide in the capsid protein GH loop relative to a corresponding parental AAV capsid protein, and wherein the variant capsid protein, when present in an AAV virion, provides for increased infectivity of the AAV virion of a retinal cell, and wherein the heterologous peptide comprises the amino acid sequence:
LAPDSTTRSA (SEQ ID NO:53).
11. An isolated, genetically modified host cell comprising the nucleic acid of claim 10 .
12. A variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises an insertion of a heterologous peptide in the GH loop of a native AAV capsid, wherein the heterologous peptide comprises the amino acid sequence:
LAPDSTTRSA (SEQ ID NO:53).