Heterologous targeting peptide grafted AAVS

Patent metadata

Field	Value
Publication number	US-11014976-B2
Application number	US-201916451049-A
Country	US
Kind code	B2
Filing date	Jun 25, 2019
Priority date	Oct 3, 2014
Publication date	May 25, 2021
Grant date	May 25, 2021

Abstract

The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associated viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same.

First claim

What is claimed is:

1. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 5, that is inserted between the first and second amino acid residues of the VP2 capsid protein; and

(b) a recombinant AAV vector comprising a transgene.

2. The rAAV of claim 1 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.

3. The rAAV of claim 2 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.

4. The rAAV of claim 3 , wherein the linker is of the formula [G] n S, wherein n is an integer in a range of 2 to 10.

5. The rAAV of claim 3 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4.

6. The rAAV of claim 5 , wherein the linker comprises SEQ ID NO: 28.

7. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 7, that is inserted between the first and second amino acid residues of the VP2 capsid protein; and

(b) a recombinant AAV vector comprising a transgene.

8. The rAAV of claim 7 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.

9. The rAAV of claim 8 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.

10. The rAAV of claim 9 , wherein the linker is of the formula [G]S, wherein n is an integer in a range of 2 to 10.

11. The rAAV of claim 9 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4.

12. The rAAV of claim 11 , wherein the linker comprises SEQ ID NO: 28.

13. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide encoded by a nucleic acid consisting of the nucleotide sequence set forth in SEQ ID NO: 29, that is inserted between the first and second amino acid residues of the VP2 capsid protein:

(b) a recombinant AAV vector comprising a transgene.

14. The rAAV of claim 13 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.

15. The rAAV of claim 14 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.

16. The rAAV of claim 15 , wherein the linker is of the formula [G]S, wherein n is an integer in a range of 2 to 10.

17. The rAAV of claim 15 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4, and wherein the linker optionally comprises SEQ ID NO: 28.

Assignees

• Univ Massachusetts

Inventors

• Esteves Miguel Sena
• Choudhury Sourav Roy

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C07K14/775 Primary

  Apolipopeptides · CPC title

• C07K14/075

  Adenoviridae · CPC title

• C12N2310/141

  MicroRNAs, miRNAs · CPC title