Adeno-associated virus variant capsids and methods of use thereof
Provided herein are variant adeno-associated virus (AAV) capsid proteins having one or more modifications in amino acid sequence relative to a parental AAV capsid protein, which, when present in an AAV virion, confer increased infectivity of one or more types of retinal cells as compared to the infectivity of the retinal cells by an AAV virion comprising the unmodified parental AAV capsid protein. Also provided are recombinant AAV virions and pharmaceutical compositions thereof comprising a variant AAV capsid protein as described herein, methods of making these rAAV capsid proteins and virions, and methods for using these rAAV capsid proteins and virions in research and in clinical practice, for example in, e.g., the delivery of nucleic acid sequences to one or more cells of the retina for the treatment of retinal disorders and diseases.
1. An infectious recombinant adeno-associated virus (rAAV) particle comprising (i) a variant AAV capsid protein comprising the amino acid sequence set forth in SEQ ID NO:42 and (ii) a heterologous nucleic acid comprising a nucleotide sequence encoding an interfering RNA with a double stranded nucleotide sequence that is complementary to the nucleotide sequence of a transcript of a rhodopsin gene associated with autosomal dominant retinitis pigmentosa, said nucleotide sequence encoding said interfering RNA operably linked to a promoter, and wherein the rAAV particle exhibits an increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV2 virion comprising a capsid protein consisting of the amino acid sequence of SEQ ID NO:2.
2. The infectious rAAV particle according to claim 1 , wherein the promoter is a rhodopsin promoter.
3. The infectious rAAV particle according to claim 1 , wherein the interfering RNA is a short hairpin RNA (shRNA).
4. A composition comprising the rAAV particle according to claim 1 and a pharmaceutically acceptable excipient.
5. The composition according to claim 4 , wherein the composition comprises from 10 8 to 10 15 vector genomes (vg) of the rAAV.
6. A method of delivering rhodopsin in one or more retinal cells of an individual in need thereof, the method comprising intravitreally or subretinally administering to the individual an rAAV comprising (i) a variant AAV capsid protein comprising the amino acid sequence set forth in SEQ ID NO:42 and (ii) a heterologous nucleic acid encoding rhodopsin (RHO), said heterologous nucleotide sequence operably linked to a promoter.