AAV virions with decreased immunoreactivity and uses therefor

Patent metadata

Field	Value
Publication number	US-9506083-B2
Application number	US-82579807-A
Country	US
Kind code	B2
Filing date	Jul 9, 2007
Priority date	Jun 19, 2003
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2.

First claim

The invention claimed is:

1. A polynucleotide encoding a mutated adeno-associated virus (AAV) VP2 capsid protein that comprises a substitution of one or more of the amino acids occurring at a position corresponding to a position of the full-length AAV-2 VP2 capsid selected from the group consisting of amino acid 132, 334, 357, 372, 377 and 395, numbered relative to the sequence of SEQ ID NO:12, wherein when the substitution is at amino acid position 357, the amino acid at position 357 is Glu, Gln or Asn, when the substitution is at amino acid position 372, the amino acid at position 372 is Phe, Lys, Asn or Gln, when the substitution is at amino acid position 334, the sequence of amino acids at amino acid positions 332-334 is Asp-Ile-Ala, when the substitution is at amino acid position 377, the sequence of amino acids at amino acid positions 376-378 is Arg-Ala-Ser, when the substitution is at amino acid position 132, the sequence of amino acid positions 132-134 is Asn-Asn-His, and when the substitution is at amino acid position 395, the sequence of amino acids at positions 394-395 is Glu-Ala,
wherein the mutated AAV VP2 capsid protein displays an increase in resistance to neutralization as compared to the wild-type (AAV) capsid.

2. The polynucleotide of claim 1 , wherein the encoded VP2 capsid protein comprises a substitution of phenylalanine, lysine, asparagine or glutamine for the naturally occurring amino acid at the position corresponding to the amino acid found at position 372 of AAV-2 VP2.

3. A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
(a) providing a recombinant AAV virion, wherein the recombinant AAV virion comprises a mutated AAV capsid protein encoded by the polynucleotide of claim 1 ; and
(b) delivering said recombinant AAV virion to said cell or tissue, whereby said protein is expressed at a level that provides a therapeutic effect.

4. The method of claim 3 , wherein said cell or tissue is a muscle cell or tissue.

5. The method of claim 4 , wherein said muscle cell or tissue is derived from skeletal muscle.

6. The method of claim 3 , wherein said recombinant AAV virion is delivered into said cell or tissue in vivo.

7. The method of claim 6 , wherein said recombinant AAV virion is delivered by intramuscular injection.

8. The method of claim 3 , wherein said recombinant AAV virion is delivered into said cell or tissue in vitro.

9. The method of claim 3 , wherein said recombinant AAV virion is delivered into the bloodstream.

10. The method of claim 9 , wherein said recombinant AAV virion is delivered intravenously.

11. The method of claim 9 , wherein said recombinant AAV virion is delivered intraarterially.

12. The method of claim 3 , wherein said recombinant AAV virion is delivered to the liver.

13. The method of claim 3 , wherein said recombinant AAV virion is delivered to the brain.

14. A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
(a) providing a recombinant AAV virion, wherein said AAV virion comprises
(i) a non-primate, mammalian adeno-associated virus (AAV) capsid protein encoded by the polynucleotide of claim 1 ; and
(ii) a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein;

(b) delivering said recombinant AAV virion to said cell or tissue, whereby said protein is expressed at a level that provides a therapeutic effect.

15. The method of claim 14 , wherein said cell or tissue is a muscle cell or tissue.

16. The method of claim 15 , wherein said muscle cell or tissue is derived from skeletal muscle.

17. The method of claim 14 , wherein said recombinant AAV virion is delivered into said cell or tissue in vivo.

18. The method of claim 17 , wherein said recombinant AAV virion is delivered by intramuscular injection.

19. The method of claim 14 , wherein said recombinant AAV virion is delivered into said cell or tissue in vitro.

20. The method of claim 14 , wherein said recombinant AAV virion is delivered into the bloodstream.

21. The method of claim 20 , wherein said recombinant AAV virion is delivered intravenously.

22. The method of claim 20 , wherein said recombinant AAV virion is delivered intraarterially.

23. The method of claim 14 , wherein said recombinant AAV virion is delivered to the liver.

24. The method of claim 14 , wherein said recombinant AAV virion is delivered to the brain.

25. The polynucleotide of claim 1 , wherein when the substitution is at amino acid position 132, the sequence of amino acid positions 132-134 is Asn-Asn-His.

26. The polynucleotide of claim 1 , wherein when the substitution is at amino acid position 334, the sequence of amino acids at amino acid positions 332-334 is Asp-Ile-Ala.

27. The polynucleotide of claim 1 , wherein when the substitution is at amino acid position 377, the sequence of amino acids at amino acid positions 376-378 is Arg-Ala-Ser.

Assignees

• Arbetman Alejandra Elena
• Colosi Peter C
• Lochrie Michael A
• Surosky Richard T
• Genzyme Corp

Inventors

• Arbetman Alejandra Elena
• Colosi Peter C
• Lochrie Michael A
• Surosky Richard T

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P3/08

  for glucose homeostasis (pancreatic hormones A61P5/48) · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P37/04

  Immunostimulants · CPC title

• A61P7/02

  Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors · CPC title