Methods and compositions for antibody-evading virus vectors
The present invention provides AAV capsid proteins comprising a modification in the amino acid sequence and virus vectors comprising the modified AAV capsid protein. The invention also provides methods of administering the virus vectors and virus capsids of the invention to a cell or to a subject in vivo.
1. A recombinant adeno associated virus (AAV) vector comprising a capsid protein and a nucleic acid, wherein the capsid protein comprises one or more of the following substitutions, wherein the amino acids are numbered according to the amino acid sequence of SEQ ID NO:1:
(a) a substitution of amino acids corresponding to amino acids 456 to 459 with the amino acid sequence SERR (SEQ ID NO:26);
(b) a substitution of amino acids corresponding to amino acids 492 to 499 with the amino acid sequence TPGGNATR (SEQ ID NO:485); and
(c) a substitution of amino acids corresponding to amino acids 588 to 597 with the amino acid sequence DLDPKATEVE (SEQ ID NO:487),
wherein the capsid protein with the substitutions of one or more of (a)-(c) has an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO:1.
2. The recombinant AAV vector of claim 1 , wherein the capsid protein with the substitutions of one or more of (a)-(c) has an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:1.
3. The recombinant AAV vector of claim 1 , wherein the capsid protein comprises at least one amino acid deletion relative to SEQ ID NO:1.
4. The recombinant AAV vector of claim 1 , wherein the capsid protein comprises at least one amino acid insertion relative to SEQ ID NO:1.
5. The recombinant AAV vector of claim 1 , wherein the one or more substitutions inhibit neutralization of infectivity of the AAV vector and/or inhibit binding of an antibody to the AAV vector, wherein the antibody binds to a capsid protein comprising the amino acid sequence of SEQ ID NO:1.
6. The recombinant AAV vector of claim 5 , wherein the antibody is a mouse monoclonal antibody selected from the group consisting of ADK1a, 4E4 and 5H7.
7. The recombinant AAV vector of claim 6 , wherein the antibody is ADK1a.
8. The recombinant AAV vector of claim 6 , wherein the antibody is 4E4.
9. The recombinant AAV vector of claim 6 , wherein the antibody is 5H7.
10. The recombinant AAV vector of claim 1 , wherein the nucleic acid encodes a heterologous polypeptide.
11. The recombinant AAV vector of claim 10 , wherein the heterologous polypeptide is a therapeutic polypeptide.
12. The recombinant AAV vector of claim 1 , wherein the nucleic acid encodes heterologous RNA sequence.
13. The recombinant virus vector of claim 12 , wherein the heterologous RNA is a functional RNA.
14. A method of producing a heterologous polypeptide in an isolated cell, comprising contacting the cell with the recombinant AAV vector of claim 10 .
15. The method of claim 14 , wherein the cell is contacted with the recombinant AAV vector in vitro, ex vivo or in vivo.
16. A method of producing a heterologous RNA in an isolated cell, comprising contacting the cell with the recombinant AAV vector of claim 12 .
17. The method of claim 16 , wherein the cell is contacted with the recombinant AAV vector in vitro, ex vivo or in vivo.
18. A pharmaceutical composition comprising the recombinant AAV vector of claim 10 , and a pharmaceutically acceptable carrier.
19. A method of producing a heterologous polypeptide in a subject, comprising administering the pharmaceutical composition of claim 18 to the subject.
20. The method of claim 19 , wherein the administration is intravenous, intraarticular, intra-lymphatic, or intra-CSF administration.
21. A pharmaceutical composition comprising the recombinant AAV vector of claim 12 , and a pharmaceutically acceptable carrier.
22. A method of producing a heterologous RNA in a subject, comprising administering the pharmaceutical composition of claim 21 to the subject.
23. The method of claim 22 , wherein the administration is intravenous, intraarticular, intra-lymphatic, or intra-CSF administration.
24. A pharmaceutical composition comprising the recombinant AAV vector of claim 1 , and a pharmaceutically acceptable carrier.