Adeno-associated virus variants and methods of use thereof
The present disclosure provides infectious recombinant adeno-associated virus (rAAV) virions that comprise a variant capsid protein and a heterologous nucleic acid. The present disclosure further provides the variant adeno-associated virus (AAV) capsid proteins (and/or a nucleic acid encoding the variant AAV capsid proteins), which confer to an infectious rAAV virion an increased resistance to human AAV neutralizing antibodies. The present disclosure further provides host cells comprising an infectious rAAV virion and/or a nucleic acid encoding a subject variant AAV capsid protein. The present disclosure further provides methods of delivering a heterologous nucleic acid to a target cell where the target cell is contacted with a subject infectious rAAV virion. The present disclosure further provides methods of delivering a gene product to an individual, the methods generally involving administering an effective amount of a subject rAAV virion to an individual in need thereof.
1. An infectious recombinant adeno-associated virus (rAAV) virion comprising:
(a) a variant adeno-associated virus (AAV) capsid protein comprising the amino acid sequence set forth in SEQ ID NO:12, wherein the rAAV virion exhibits greater resistance to neutralization by a neutralizing antibody compared to AAV2; and
(b) a heterologous nucleic acid comprising a nucleotide sequence encoding a heterologous gene product.
2. The infectious rAAV of claim 1 , wherein the variant AAV capsid protein consists of the amino acid sequence set forth in SEQ ID NO: 12.
3. The infectious rAAV of claim 1 , wherein the rAAV exhibits increased transduction of mammalian cells in the presence of neutralizing antibodies compared to the transduction of mammalian cells exhibited by AAV serotype 2 (AAV2).
4. The infectious rAAV of claim 3 , wherein the mammalian cells are liver cells, pancreatic cells, skeletal muscle cells, heart muscle cells, fibroblasts, retinal cells, synovial joint cells, lung cells, T cells, neurons, glial cells, stem cells, endothelial cells, or cancer cells.
5. The infectious rAAV of claim 4 , wherein the stem cells are hematopoietic stem cells, hematopoietic progenitor cells, neural stem cells, neural progenitor cells, neural crest stem cells, embryonic stem cells, induced pluripotent stem cells (iPS cells), mesenchymal stem cells, mesodermal stem cells, liver stem cells, pancreatic stem cells, pancreatic progenitor cells, muscle stem cells, or retinal stem cells.
6. The infectious rAAV of claim 1 , wherein the heterologous nucleic acid comprises a nucleotide sequence encoding an RNA interfering agent.
7. The infectious rAAV of claim 1 , wherein the heterologous nucleic acid comprises a nucleotide sequence encoding a polypeptide.
8. A pharmaceutical composition comprising:
a) the rAAV virion according to claim 1 ; and
b) a pharmaceutically acceptable excipient.
9. The pharmaceutical composition of claim 8 , comprising from 10 6 to 10 15 rAAV virions.