Adeno-associated virus (AAV) glades, sequences, vectors containing same, and uses therefor

Patent metadata

Field	Value
Publication number	US-9737618-B2
Application number	US-201514803683-A
Country	US
Kind code	B2
Filing date	Jul 20, 2015
Priority date	Sep 30, 2003
Publication date	Aug 22, 2017
Grant date	Aug 22, 2017

Abstract

Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided.

First claim

The invention claimed is:

1. A recombinant adeno-associated virus (AAV) comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp1 having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.

2. The recombinant AAV according to claim 1 wherein the AAV ITR is from AAV2.

3. The recombinant AAV according to claim 1 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).

4. The recombinant AAV according to claim 1 , wherein the gene product is an antibody.

5. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp1 having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.

6. A recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp3 having the amino acid sequence of amino acids 204 to 738 of SEQ ID NO: 88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.

7. The recombinant AAV according to claim 6 wherein the AAV ITR is from AAV2.

8. The recombinant AAV according to claim 6 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).

9. The recombinant AAV according to claim 6 , wherein the gene product is an antibody.

10. The recombinant AAV according to claim 6 , wherein the vp1 protein has the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88.

11. The recombinant AAV according to claim 6 , wherein the vp2 protein has the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88.

12. The recombinant AAV according to claim 6 , wherein the vp1 protein has the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, the vp2 protein has the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88 and the vp3 protein has the amino acid sequence of amino acids 204 to 738 of SEQ ID NO:88.

13. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp2 having the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.

14. A recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp2 having the amino acid sequence of amino acids 138 to 738 of SEQ ID NO: 88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression thereof in a host cell.

15. The recombinant AAV according to claim 14 wherein the AAV ITR is from AAV2.

16. The recombinant AAV according to claim 14 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).

17. The recombinant AAV according to claim 14 , wherein the gene product is an antibody.

18. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp3 having the amino acid sequence of amino acids 204 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.

19. A method of generating a recombinant AAV comprising an AAV hu.37 capsid comprising the steps of culturing a host cell containing: (a) a nucleic acid sequence encoding the AAVhu.37 vp1 capsid protein having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO: 88; (b) a sequence encoding an AAV rep; (c) a nucleic acid sequence comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell and (d) sufficient helper functions for packaging the AAV.

Assignees

• Univ Pennsylvania

Inventors

• Wilson James M
• Gao Guangping
• Alvira Mauricio R
• Vandenberghe Luk H

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C07K14/005

  from viruses · CPC title

• C12N2750/14122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C12N2750/14142

  virus or viral particle as vehicle, e.g. encapsulating small organic molecule · CPC title

• C07K14/705

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title