Search

US-20260124322-A1 - ATP7B GENE THERAPY

US20260124322A1US 20260124322 A1US20260124322 A1US 20260124322A1US-20260124322-A1

Abstract

Provided herein are improved expression constructs for the expression of ATP7B, vectors, and pharmaceutical compositions comprising such constructs. Also provided are methods of treating disease, including, but not limited to, Wilson's Disease.

Inventors

  • Alexandria Forbes
  • Josefa SULLIVAN
  • Edgar Hernandez
  • Matthew During
  • Ce Feng LIU

Assignees

  • MEIRAGTX UK II LIMITED

Dates

Publication Date
20260507
Application Date
20231011

Claims (20)

  1. 1 . An expression construct comprising: (a) a promoter; (b) a sequence encoding (ATPase Copper Transporting Beta) ATP7B, operatively linked to the promoter; and (c) a polyadenylation signal.
  2. 2 . The expression construct of claim 1 , wherein the promoter comprises a sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 4-33.
  3. 3 . The expression construct of claim 2 , wherein the promoter comprises a sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 4-33.
  4. 4 . The expression construct of claim 3 , wherein the promoter comprises a sequence that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 4-33.
  5. 5 . The expression construct of claim 4 , wherein the promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 4-33.
  6. 6 . The expression construct of claim 1 , wherein the promoter comprises a sequence that is at least 80% identical to any one of SEQ ID NO:4, SEQ ID NO: 10, or SEQ ID NO:11.
  7. 7 . The expression construct of claim 6 , wherein the promoter comprises a sequence that is at least 90% identical to any one of SEQ ID NO:4, SEQ ID NO: 10, or SEQ ID NO:11.
  8. 8 . The expression construct of claim 7 , wherein the promoter comprises a sequence that is at least 95% identical to any one of SEQ ID NO:4, SEQ ID NO:10, or SEQ ID NO: 11.
  9. 9 . The expression construct of claim 8 , wherein the promoter comprises any one of SEQ ID NO: 4, SEQ ID NO: 10, or SEQ ID NO:11.
  10. 10 . The expression construct of any one of claims 1-8 , wherein the sequence encoding ATP7B is codon-optimized.
  11. 11 . The expression construct of any one of claims 1-8 , wherein the sequence encoding ATP7B comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 35-48.
  12. 12 . The expression construct of claim 11 , wherein the sequence encoding ATP7B comprises a sequence that is at least 90% identical to any one of SEQ ID NOs: 35-48.
  13. 13 . The expression construct of claim 12 , wherein the sequence encoding ATP7B comprises a sequence that is at least 95% identical to any one of SEQ ID NOs: 35-48.
  14. 14 . The expression construct of claim 13 , wherein the sequence encoding ATP7B comprises a sequence selected from the group consisting of SEQ ID NOs: 35-48.
  15. 15 . The expression construct of any one of claims 1-8 , wherein the sequence encoding ATP7B comprises a sequence that is at least 80% identical to SEQ ID NO:39 or SEQ ID NO:41.
  16. 16 . The expression construct of claim 15 , wherein the sequence encoding ATP7B comprises a sequence that is at least 90% identical to SEQ ID NO:39 or SEQ ID NO:41.
  17. 17 . The expression construct of claim 16 , wherein the sequence encoding ATP7B comprises a sequence that is at least 95% identical to SEQ ID NO:39 or SEQ ID NO:41.
  18. 18 . The expression construct of claim 17 , wherein the sequence encoding ATP7B comprises SEQ ID NO:39 or SEQ ID NO:41.
  19. 19 . The expression construct of any of the preceding claims , wherein the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 80% identical to any one of SEQ ID NOs: 118-128.
  20. 20 . The expression construct of claim 19 , wherein the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 90% identical to any one of SEQ ID NOs: 118-128.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of the earlier filing date of U.S. Provisional Patent No. 63/379,113, filed Oct. 11, 2022, which is hereby incorporated by reference in its entirety. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (SeqList-162027.53176.xml; Size: 479,210 bytes; and Date of Creation: Oct. 10, 2023) is herein incorporated by reference in its entirety. FIELD The present disclosure relates generally to the field of molecular biology and medicine. More particularly, the methods and compositions herein are useful for treating Wilson's Disease. BACKGROUND Wilson's Disease (WD) is caused by autosomal recessive, loss-of-function mutations in the ATPase copper transporting beta (ATP7B) gene which lead to pathological accumulation of copper in the liver, brain, and other tissues. Symptoms of WD include Parkinson's Disease like neurological defects (including dystonia/bradykinesia) and hepatologic defects associated with cirrhosis. The prevalence of Wilson's Disease is 1 in ˜30,000 people. ATP7B is a transmembrane copper ion transporter. When the cellular copper concentration increases, ATP7B translocates to the lysosome and pumps copper into vesicles so that the copper can be excreted via the bile duct in the liver. Due it is large size (1465 amino acids), ATP7B is too large to be effectively packaged into adeno-associated virus (AAV), making ATP7B gene therapy difficult. Accordingly, enhanced ATP7B expression constructs for the treatment of Wilson's Disease are urgently needed. SUMMARY Provided herein are improved expression constructs for the expression of ATP7B, vectors, and pharmaceutical compositions comprising such constructs, and methods of using such constructs, vectors, and pharmaceutical compositions. In one aspect, provided is an expression construct comprising: (a) a promoter;(b) a sequence encoding (ATPase Copper Transporting Beta) ATP7B, operatively linked to the promoter; and(c) a polyadenylation signal. In some embodiments, the promoter comprises a sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs:4-33. In some embodiments, the promoter comprises a sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs:4-33. In some embodiments, the promoter comprises a sequence that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs:4-33. In some embodiments, the promoter comprises a sequence selected from the group consisting of SEQ ID NOs:4-33. In some embodiments, the promoter comprises a sequence that is at least 80% identical to any one of SEQ ID NO:4, SEQ ID NO:10, or SEQ ID NO:11. In some embodiments, the promoter comprises a sequence that is at least 90% identical to any one of SEQ ID NOs:4, 10, or 11. In some embodiments, the promoter comprises a sequence that is at least 95% identical to any one of SEQ ID NOs:4, 10, or 11. In some embodiments, the promoter comprises any one of SEQ ID NOs:4, 10, or 11. In some embodiments, the sequence encoding ATP7B is codon-optimized. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 80% identical to any one of SEQ ID NOs:35-48. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 90% identical to any one of SEQ ID NOs:35-48. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 95% identical to any one of SEQ ID NOs:35-48. In some embodiments, the sequence encoding ATP7B comprises a sequence selected from the group consisting of SEQ ID NOs:35-48. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 80% identical to SEQ ID NO:39 or SEQ ID NO:41. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 90% identical to SEQ ID NO:39 or SEQ ID NO:41. In some embodiments, the sequence encoding ATP7B comprises a sequence that is at least 95% identical to SEQ ID NO:39 or SEQ ID NO:41. In some embodiments, the sequence encoding ATP7B comprises SEQ ID NO:39 or SEQ ID NO:41. In some embodiments, the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 80% identical to any one of SEQ ID NOs:118-128. In some embodiments, the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 90% identical to any one of SEQ ID NOs:118-128. In some embodiments, the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 95% identical to any one of SEQ ID NOs:118-128. In some embodiments, the sequence encoding ATP7B encodes a protein comprising any one of SEQ ID NOs:118-128. In some embodiments, the sequence encoding ATP7B encodes a protein comprising a sequence that is at least 80% identical to SEQ ID NO:118 or SEQ ID NO:123. In some embodiments, the sequence encoding ATP7B encodes a protein co