Adeno-associated virus virions with variant capsid and methods of use thereof

Patent metadata

Field	Value
Publication number	US-2016376323-A1
Application number	US-201615244884-A
Country	US
Kind code	A1
Filing date	Aug 23, 2016
Priority date	Apr 22, 2011
Publication date	Dec 29, 2016
Grant date	—

Abstract

The present disclosure provides adeno-associated virus (AAV) virions with altered capsid protein, where the AAV virions exhibit greater infectivity of retinal cells, when administered via intravitreal injection, compared to wild-type AAV. The present disclosure further provides methods of delivering a gene product to a retinal cell in an individual, and methods of treating ocular disease.

First claim

1 - 27 . (canceled)

28 . A recombinant adeno-associated virus (rAAV) virion comprising:
a) a variant AAV capsid protein, wherein the variant AAV capsid protein comprises a peptide insertion relative to a corresponding parental AAV capsid protein, wherein the peptide insertion comprises an amino acid sequence selected from the group consisting of NETITRP (SEQ ID NO: 14), KAGQANN (SEQ ID NO: 15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO: 57), RAGGSVG (SEQ ID NO: 58), AVDTTKF (SEQ ID NO: 59), and STGKVPN (SEQ ID NO: 60), wherein the insertion site is located between two adjacent amino acids at a position between amino acids corresponding to amino acids 570 and 611 of VP1 of AAV2 or the corresponding position in the capsid protein of another AAV serotype; and b) a heterologous nucleic acid comprising a nucleotide sequence encoding a gene product.

29 . The rAAV virion of claim 28 , wherein the variant capsid protein confers increased infectivity of a retinal cell by the rAAV virion compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.

30 . The rAAV virion of claim 29 , wherein the retinal cell is a photoreceptor, a retinal ganglion cell, a Müller cell, a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelium cell.

31 . The rAAV virion of claim 28 , wherein the insertion site is located between amino acids corresponding to amino acids 587 and 588 of VP1 of AAV2 or the corresponding position in the capsid protein of another AAV serotype.

32 . The rAAV virion of claim 28 , wherein the rAAV virion exhibits at least 10-fold increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.

33 . The rAAV virion of claim 28 , wherein the rAAV virion exhibits at least 50-fold increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.

34 . The rAAV virion of claim 28 , wherein the gene product is an interfering RNA or an aptamer.

35 . The rAAV virion of claim 28 , wherein the gene product is a polypeptide.

36 . The rAAV virion of claim 35 , wherein the polypeptide is a neuroprotective polypeptide, an anti-angiogenic polypeptide, or a polypeptide that enhances function of a retinal cell.

37 . The rAAV virion of claim 28 , wherein the peptide insertion comprises an amino acid sequence selected from the group consisting of NETITRP (SEQ ID NO: 14), KAGQANN (SEQ ID NO: 15), and KDPKTTN (SEQ ID NO: 16).

38 . The rAAV virion of claim 28 , wherein the parental capsid protein is an AAV2 capsid protein and the insertion site is between amino acids 570 and 611 of the parental AAV2 VP1 capsid protein.

39 . A pharmaceutical composition comprising:
a) a recombinant adeno-associated virus virion of claim 28 ; and b) a pharmaceutically acceptable excipient.

40 . A method of delivering a gene product to a retinal cell in an individual, the method comprising administering to the individual a recombinant adeno-associated virus (rAAV) virion according to claim 28 .

41 . The method of claim 40 , wherein the gene product is an interfering RNA or an aptamer.

42 . The method of claim 40 , wherein the gene product is a polypeptide.

43 . The method of claim 42 , wherein the polypeptide is a neuroprotective factor, an anti-angiogenic polypeptide, an anti-apoptotic factor, or a polypeptide that enhances function of a retinal cell.

44 . The method of claim 42 , wherein the polypeptide is glial derived neurotrophic factor, fibroblast growth factor 2, neurturin, ciliary neurotrophic factor, nerve growth factor, brain derived neurotrophic factor, epidermal growth factor, rhodopsin, X-linked inhibitor of apoptosis, retinoschisin, RPE65, retinitis pigmentosa GTPase-interacting protein-1, peripherin, peripherin-2, a rhodopsin, or Sonic hedgehog.

45 . A method of treating an ocular disease, the method comprising administering to an individual in need thereof an effective amount of a recombinant adeno-associated virus (rAAV) virion according to claim 28 .

46 . The method of claim 45 , wherein said administering is by intraocular injection.

47 . The method of claim 45 , wherein said administering is by intravitreal injection.

48 . The method of claim 45 , wherein the ocular disease is glaucoma, retinitis pigmentosa, macular degeneration, retinoschisis, Leber's Congenital Amaurosis, diabetic retinopathy, achromotopsia, or color blindness.

49 . An isolated nucleic acid comprising a nucleotide sequence that encodes a variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises a peptide insertion relative to a corresponding parental AAV capsid protein, wherein the peptide insertion comprises an amino acid sequence selected from the group consisting of NETITRP (SEQ ID NO: 14), KAGQANN (SEQ ID NO: 15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO: 57), RAGGSVG (SEQ ID NO: 58), AVDTTKF (SEQ ID NO: 59), and STGKVPN (SEQ ID NO: 60), wherein the insertion site is located between two adjacent amino acids at a position between amino acids corresponding to amino acids 570 and 611 of VP1 of AAV2 or the corresponding position in the capsid protein of another AAV serotype.

50 . The isolated nucleic acid of claim 49 , wherein the variant AAV capsid protein, when present in an AAV virion, provides for increased infectivity of a retinal cell by the AAV virion compared to the infectivity of a retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.

51 . The isolated nucleic acid of claim 50 , wherein the retinal cell is a photoreceptor, a retinal ganglion cell, a Müller cell, a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelium cell.

52 . The isolated nucleic acid of claim 49 , wherein the insertion site is located between amino acids corresponding to amino acids 587 and 588 of VP1 of AAV2 or the corresponding position in the capsid protein of another AAV serotype.

53 . The isolated nucleic acid of claim 49 , wherein the parental capsid protein is an AAV2 capsid protein and the insertion site is between amino acids 570 and 611 of the parental AAV2 VP1 capsid protein.

54 . An isolated, genetically modified host cell comprising the nucleic acid of claim 49 .

55 . A variant adeno-associated virus (AAV) capsid protein, wherein the variant AAV capsid protein comprises a peptide insertion relative to a corresponding parental AAV capsid protein, wherein the peptide insertion comprises an amino acid sequence selected from the group consisting of NETITRP (SEQ ID NO: 14), KAGQANN (SEQ ID NO: 15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO: 57), RAGGSVG (SEQ ID NO: 58), AVDTTKF (SEQ ID NO: 59), and STGKVPN (SEQ ID NO: 60), wherein the insertion site is located between two adjacent amino acids at a position between amino acids corresponding to amino acids 570 and 611 of VP1 of AAV2 or the corresponding position in the capsid protein of another AAV serotype.

56 . The variant AAV capsid protein of claim 55 , wherein the variant capsid protein, when present in an AAV virion, confers increased infectivity of a retinal cell by the AAV virion compared to the infectivity of t

Assignees

• Univ California

Inventors

• Schaffer David V
• Klimczak Ryan R
• Koerber James T
• Flannery John G
• Dalkara Mourot Deniz
• Visel Meike
• Byrne Leah C T

Classifications

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P43/00

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

• A61P9/00

  Drugs for disorders of the cardiovascular system · 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

• A61P27/02

  Ophthalmic agents · CPC title