Methods and compositions for modulating the interaction between adeno-associated virus (aav) and the aav receptor (aavr) for altered bio-distribution of aav

Patent metadata

Field	Value
Publication number	US-2024060087-A1
Application number	US-202218271250-A
Country	US
Kind code	A1
Filing date	Jan 7, 2022
Priority date	Jan 8, 2021
Publication date	Feb 22, 2024
Grant date	—

Abstract

This disclosure describes compositions and methods for altering the bio-distribution of adeno-associated viruses (AAVs) in subjects.

First claim

What is claimed is:

1 . A virus comprising a non-naturally occurring, modified AAV VP1 capsid protein, comprising
an amino acid sequence having at least 95% sequence identity to an amino acid sequence of an unmodified AAV VP1 capsid protein when the amino acid sequence of the modified AAV capsid protein and the amino acid sequence of the unmodified AAV VP1 capsid protein are aligned using a basic local alignment search tool (BLAST) program with default algorithm parameters; wherein the amino acid sequence of the modified VP1 capsid protein differs from the amino acid sequence of the unmodified VP1 capsid protein in at least one amino acid position selected from the group consisting of amino acid positions Q263, S264, G265, A266, S267, N268, H271, N382, G383, S384, Q385, S446, R471, W502, T503, D528, D529, Q589, K706, and V708, which are numbered relative to AAV2 VP1 capsid protein (SEQ ID NO:1) when SEQ ID NO:1 and the amino acid sequence of the unmodified AAV capsid protein are aligned using the basic local alignment search tool (BLAST) program with default algorithm parameters.

2 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises at least one amino acid residue selected from the group consisting of 446R, 471A, and 708T.

3 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises at least two amino acid residues selected from the group consisting of 446R, 471A, and 708T.

4 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises amino acid residues 446R, 471A, and 708T.

5 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises at least one amino acid residue selected from the group consisting of 446S, 471S, and 708A.

6 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises at least two amino acid residues selected from the group consisting of 446S, 471S, and 708A.

7 . The virus of claim 1 , wherein the amino acid sequence of the modified VP1 capsid protein comprises 446S, 471S, and 708A.

8 . The virus of any one of claims 1 to 7 , wherein the amino acid sequence of the modified VP1 capsid protein differs from the amino acid sequence of the unmodified VP1 capsid protein only in one or more of the amino acid positions Q263, S264, G265, A266, S267, N268, H271, N382, G383, S384, Q385, S446, R471, W502, T503, D528, D529, Q589, K706, and V708, and in no other amino acid positions in the unmodified VP1 capsid protein.

9 . The virus of any one of claims 1 to 8 , wherein the unmodified VP1 capsid protein is selected from the group consisting of a VP1 capsid protein from AAV1, AAV2, AAV3, AAV6, AAV7, AAV8, AAV9, rh.10, hu.37, LK-03, AAV5, AAV10, Hu68; Anc80; Anc81; Anc82; Anc83; Anc84; Anc94; Anc113; Anc126; Anc127; Anc80L27; Anc80L59; Anc80L60; Anc80L62; Anc80L65; Anc80L33; Anc80L36; Anc80L44; Anc80L1; Anc110; Anc80DI; AAV1 vp1; AAV2 vp1; AAV9vp1; Anc80; Anc126; Anc127; AAV3; AAV7; AAV8; rh10; hu37; and hu.68.

10 . The virus of any one of claims 1 to 9 , wherein the non-naturally occurring, modified AAV VP1 capsid protein comprises an amino acid sequence having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of the unmodified AAV VP1 capsid protein when the amino acid sequence of the modified AAV capsid protein and the amino acid sequence of the unmodified AAV VP1 capsid protein are aligned using a basic local alignment search tool (BLAST) program with default algorithm parameters.

11 . A modified, assembly-competent recombinant AAV (rAAV), comprising:
VP1, VP2, and VP3 capsid proteins, and a recombinant nucleic acid vector, wherein the VP1 capsid protein is the modified VP1 capsid protein of any one of claims 1 to 10 .

12 . A modified, assembly-competent recombinant AAV (rAAV), comprising:
VP1, VP2, and VP3 capsid proteins; and a recombinant nucleic acid vector, wherein at least the VP1 capsid protein is a non-naturally occurring, modified VP1 capsid protein comprising an amino acid sequence having at least 95% sequence identity to an amino acid sequence of an unmodified AAV VP1 capsid protein when the amino acid sequence of the modified AAV capsid protein and the amino acid sequence of the unmodified AAV VP1 capsid protein are aligned using a basic local alignment search tool (BLAST) program with default algorithm parameters, and wherein the modified VP1 capsid protein differs from the unmodified VP1 capsid protein in comprising a means for altering biodistribution of the modified rAAV following administration of the modified rAAV to a first mammalian subject as compared to biodistribution of an unmodified rAAV following administration of the unmodified rAAV having the unmodified VP1 capsid protein to a second mammalian subject of the same type as the first mammalian subject, wherein the unmodified rAAV comprises VP1, VP2, and VP3 capsid proteins having amino acid sequences identical to those of the modified rAAV except for said means.

13 . The modified rAAV of claim 11 or claim 12 , wherein the modified rAAV achieves higher transduction of liver cells following administration to a first mammalian subject as compared to transduction of liver cells following administration of the unmodified rAAV comprising the unmodified VP1 capsid protein to a second mammalian subject of the same type as the first mammalian subject.

14 . The modified rAAV of any one of claims 11 to 13 , wherein the modified rAAV exhibits higher expression in liver cells of an expressible polypeptide encoded by the recombinant nucleic acid vector following administration to a first mammalian subject as compared to expression in liver cells of the expressible polypeptide following administration of an unmodified rAAV comprising the unmodified VP1 capsid protein to a second mammalian subject of the same type as the first mammalian subject.

15 . The modified rAAV of claim 11 or claim 12 , wherein the modified rAAV achieves lower transduction of liver cells following administration to a first mammalian subject as compared to transduction of liver cells following administration of an unmodified rAAV comprising the unmodified VP1 capsid protein to a second mammalian subject of the same type as the first mammalian subject.

16 . The modified rAAV of any one of claim 11 , 12 , or 15 , wherein the modified rAAV exhibits lower expression in liver cells of an expressible polypeptide encoded by the recombinant nucleic acid vector following administration to a first mammalian subject as compared to expression in liver cells of the expressible polypeptide following administration of an unmodified rAAV comprising the unmodified VP1 capsid protein to a second mammalian subject of the same type as the first mammalian subject.

17 . The modified rAAV of any one of claims 11 to 16 , wherein the modified rAAV has an altered interaction with an AAV receptor (AAVR) expressed on liver cells of the first mammalian subject as compared to an unmodified rAAV comprising the unmodified VP1 capsid protein with an AAVR expressed on liver cells of the second mammalian subject.

18 . The modified rAAV of claim 17 , wherein the modified rAAV has increased interaction with an AAV receptor (AAVR) expressed on liver cells of the first mammalian subject as compared to an unmodified rAAV comprising the unmodified VP1 capsid protein with an AAVR expressed on liver cells of the second mammalian subject.

19 . The modifi

Assignees

• Massachusetts Eye & Ear Infirmary
• Schepens Eye Res Inst

Inventors

• Zinn Eric
• Tipper Christopher
• Vandenberghe Luk H

Classifications

• C12N15/86 Primary

  Viral vectors · CPC title

• C07K14/005 Primary

  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

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2750/14145

  Special targeting system for viral vectors · CPC title