Adeno-associated virus virions with variant capsid
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. A recombinant adeno-associated virus 2 (rAAV2) virion comprising:
a) a variant AAV capsid protein, wherein the variant AAV capsid protein comprises a peptide insertion relative to a corresponding parental AAV capsid protein as set forth in SEQ ID NO: 1, wherein the peptide insertion is between amino acids 587 and 588 relative to SEQ ID NO: 1, and wherein the peptide insertion comprises the amino acid LALGETTRPA (SEQ ID NO: 45); and
b) a heterologous nucleic acid comprising a nucleotide sequence encoding an anti-angiogenic polypeptide.
2. The rAAV2 virion of claim 1 , wherein the variant AAV capsid protein comprises: i) an amino acid sequence having at least 98% sequence identity to SEQ ID NO:1; and ii) the peptide insertion.
3. The rAAV2 virion of claim 1 , wherein the variant AAV capsid protein comprises: i) an amino acid sequence as set forth in SEQ ID NO: 1; and ii) the peptide insertion.
4. The rAAV2 virion of claim 1 , wherein the anti-angiogenic polypeptide is a soluble vascular endothelial growth factor (VEGF) receptor, a VEGF-binding antibody, an Fc fusion polypeptide comprising a soluble Flt polypeptide.
5. The rAAV2 virion of claim 1 , wherein the anti-angiogenic polypeptide is an Fc fusion polypeptide comprising a soluble Flt polypeptide.
6. The rAAV2 virion of claim 2 , wherein the anti-angiogenic polypeptide is an Fc fusion polypeptide comprising a soluble Flt polypeptide.
7. The rAAV2 virion of claim 3 , wherein the anti-angiogenic polypeptide is an Fc fusion polypeptide comprising a soluble Flt polypeptide.
8. A pharmaceutical composition comprising:
a) a recombinant adeno-associated virus 2 virion of claim 6 ; and
b) a pharmaceutically acceptable excipient.