JP-2022515338-A5 -

Dates

Publication Date

20221212

Application Date

20191204

Description

This disclosure generally relates to the field of gene therapy, and more specifically to recombinant adeno-associated virus (AAV) vector particles having novel capsid proteins (also known as AAV viral vectors), their manufacture, and their use for delivering transgenes to treat or prevent disease or disorder. In certain embodiments, for example, the following are provided: (Item 1) A nucleic acid encoding an AAV capsid protein comprising a VP1 portion, a VP2 portion, and a VP3 portion, wherein the VP3 portion includes variable regions (VR) I to IX, (a) VR-II contains the amino acid sequence DNNGVK (SEQ ID NO: 54), (b) VR-III contains the amino acid sequence NDGS (SEQ ID NO: 55), (c) VR-IV contains the amino acid sequence INGSGQNQQT (SEQ ID NO: 56), (d) VR-V contains the amino acid sequence RVSTTTGQNNNSNFAWTA (SEQ ID NO: 57), (e) VR-VI contains the amino acid sequence HKEGEDRFPPLSG (SEQ ID NO: 58), (f) VR-VII contains the amino acid sequence KQNAARDNADYSDV (SEQ ID NO: 59), (g) VR-VII contains the amino acid sequence ADNLQQQNTAPQI (SEQ ID NO: 60), and (h) VR-IX is a nucleic acid containing the amino acid sequence NYYKSTSVDF (SEQ ID NO: 61). (Item 2) The nucleic acid according to item 1, wherein the VR-I region includes NSTSGGSS (SEQ ID NO: 53) or SSTSGSS (SEQ ID NO: 87). (Item 3) The nucleic acid according to item 1 or 2, wherein the VR-I region includes SASTGAS (SEQ ID NO: 52). (Item 4) The nucleic acid according to any one of items 1 to 3, wherein the VP3 portion has the sequence of sequence number 41. (Item 5) The nucleic acid according to item 2, wherein the encoded AAV capsid amino acid sequence is at least 95% identical to the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 84. (Item 6) The nucleic acid according to item 3, wherein the encoded AAV capsid amino acid sequence is at least 95% identical to the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 34. (Item 7) The nucleic acid according to item 4, wherein the nucleic acid sequence is at least 95% identical to a nucleotide sequence selected from sequence numbers 18 to 23. (Item 8) The nucleic acid according to item 7, wherein the nucleic acid sequence is 100% identical to a nucleotide sequence selected from sequence numbers 18 to 23. (Item 9) A vector containing nucleic acids as described in items 1 to 8. (Item 10) AAV capsid proteins encoded by the nucleic acids listed in items 1-8. (Item 11) The AAV capsid protein according to item 10, wherein the protein comprises the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 34. (Item 12) An AAV viral vector comprising the AAV capsid protein encoded by the nucleic acids described in items 1 to 8, and an AAV vector, wherein the AAV vector is oriented in the 5' to 3' direction, (a) First AAV inverse terminal repeat, (b) promoter, (c) a heterologous nucleic acid; (d) Poly A tail, and (e) An AAV viral vector containing a second AAV reverse-terminal repeat. (Item 13) The AAV virus vector according to item 12, wherein the heterogeneous nucleic acid is operably linked to a constitutive promoter. (Item 14) The AAV virus vector according to item 12, wherein the heterogeneous nucleic acid encodes a polypeptide. (Item 15) An AAV virus vector as described in item 12, wherein the heterogeneous nucleic acid encodes antisense RNA, microRNA, or RNAi. (Item 16) The AAV virus vector according to item 12, wherein the AAV capsid protein comprises the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 34. (Item 17) (i) an AAV capsid protein having the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 34, (ii) AAV vector, wherein the AAV vector is in the direction from 5' to 3', (a) First AAV inverse terminal repeat, (b) promoter, (c) a heterologous nucleic acid; (d) Poly A tail, and (e) An AAV viral vector comprising a second AAV reverse-terminal repeat. (Item 18) The AAV virus vector according to item 12 or 17, wherein the heterogeneous nucleic acid encodes mRNA, siRNA, gRNA, or microRNA. (Item 19) The AAV virus vector according to item 12 or 17, wherein the heterogeneous nucleic acid encodes a polypeptide. (Item 20) The AAV virus vector described in item 19, wherein the heterologous gene sequence encodes cystic fibrosis transmembrane conductance regulator (CFTR), CLN3 protein, alpha-galactosidase A (GLA), or acid alpha-glucosidase (GAA). (Item 21) An AAV virus vector as described in item 20, in which a heterologous sequence encodes CFTR. (Item 22) The AAV virus vector according to item 21, wherein the CFTR comprises the amino acid sequence encoded by sequence number 4. (Item 23) The AAV virus vector according to item 19, wherein the heterologous gene sequence encodes a protein having an amino acid sequence that is at least 70%, 80%, 90%, or 99% identical to any one of sequence numbers 5, 8, 11, and 14. (Item 24) The AAV vir