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. A method of treating a human subject with choroideremia, the method comprising administering to the subject by intravitreal injection, a pharmaceutical composition comprising a therapeutically effective amount of an infectious recombinant adeno-associated virus (rAAV) virion comprising (i) a variant AAV capsid protein comprising the amino acid sequence as set forth in SEQ ID NO: 42 and (ii) a heterologous nucleic acid comprising a nucleotide sequence encoding a Rab escort protein-1 (REP 1), said nucleotide sequence operably linked to an ubiquitous promoter that directs expression of the REP1 in one or more retinal pigmented epithelial cells and in one or more rod photoreceptor cells, thereby treating said human subject with choroideremia.
2. The method according to claim 1 , wherein the ubiquitous promoter is a CAG promoter.
3. The method according to claim 1 , comprising administering to the subject a pharmaceutical composition comprising from 10 11 to 10 15 vg particles of said rAAV and a pharmaceutically acceptable excipient.
4. A method of treating a human subject with choroideremia, the method comprising
administering to the human subject by subretinal injection, a pharmaceutical composition comprising a therapeutically effective amount of an infectious recombinant adeno-associated virus (rAAV) virion comprising (i) a variant AAV capsid protein comprising the amino acid sequence as set forth in SEQ ID NO: 42 and (ii) a heterologous nucleic acid comprising a nucleotide sequence encoding a Rab escort protein-1 (REP 1), said nucleotide sequence operably linked to an ubiquitous promoter that directs expression of the REP1 in one or more retinal pigmented epithelial cells and in one or more rod photoreceptor cells, thereby treating said human subject with choroideremia.
5. The method according to claim 4 , wherein the ubiquitous promoter is a CAG promoter.
6. The method according to claim 4 , comprising administering to the subject a pharmaceutical composition comprising from 10 11 to 10 15 vg particles of said rAAV and a pharmaceutically acceptable excipient.