Methods and compositions for treating Huntington's disease

• US 9,499,597 B2
• Filed: 07/29/2014
• Issued: 11/22/2016
• Est. Priority Date: 02/29/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Inventors: Miller, Jeffrey C., Rebar, Edward J., Zhang, H. Steve
• Primary Examiner(s): Weidner, Adam M
• Application Number:
  US14/446,189
• Publication Number:
  US 20140336133A1
• Time in Patent Office: 847 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4703 Inhibitors; Suppressors
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C07K 2319/71 containing domain for transcriptional activaation, e.g. VP16
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/907 in mammalian cells

Abstract

Disclosed herein are compositions comprising non-naturally occurring zinc finger domains, fusion proteins comprising these zinc finger domains, polynucleotides encoding these proteins, cells expressing these proteins and pharmaceutical compositions comprising these proteins or polynucleotides as well as methods of modifying an Htt gene using these compositions for treating or preventing Huntington'"'"'s Disease.

Claims

1. A non-naturally occurring zinc finger protein consisting of 4, 5 or 6 zinc finger domains ordered F1 to F4, F1 to F5 or F1 to F6, respectively, wherein the zinc finger protein binds to an Htt gene and comprises one- or two-finger domains at positions F2 or F2 and F3, the two-finger domains comprising the recognition helix regions sequences in F2 and F3 as shown in a single row of Table 2C and the one-finger domain of F2 comprising MACCRYA (SEQ ID NO:

175);

RSANLRE (SEQ ID NO;

     176);

RNADRKK (SEQ ID NO;

177);

RSDVLSE SEQ ID NO;

42);

RSAVLSE (SEQ ID NO;

148);

QSGDLTR (SEQ ID NO;

18);

QNATRIK (SEQ ID NO;

178) or QSSDLSR (SEQ ID NO;

31).

2. The zinc finger protein of claim 1, wherein the zinc finger protein binds entirely or partially outside the CAG repeat region of the Htt gene.

3. The zinc finger protein of claim 1, wherein the zinc finger protein binds to sequences within the CAG repeat region of the Htt gene.

4. The zinc finger protein of claim 1, further comprising a dimerization domain that allows multimerization of zinc finger proteins when bound to DNA.

5. A host cell comprising one or more zinc finger proteins of claim 1.

6. A pharmaceutical composition comprising one or more zinc finger proteins according to claim 1.

7. A polynucleotide encoding one or more zinc finger proteins of claim 1.

8. A pharmaceutical composition comprising one or more polynucleotides according to claim 7.

9. A fusion protein comprising a zinc finger protein of claim 1 and a functional domain, wherein the functional domain is selected from the group consisting of a transcriptional activation domain, a transcriptional repression domain, and a nuclease domain.

10. A method of treating Huntington'"'"'s Disease, the method comprising administering one or more polynucleotides encoding one or more fusion proteins according to claim 9 to a subject in need thereof.

11. A method of modifying expression of an Htt gene in a cell, the method comprising administering to the cell one or more polynucleotides encoding one or more fusion proteins according to claim 9.

12. The method of claim 11, wherein the Htt gene comprises at least one mutant allele.

13. The method of claim 11, wherein the Htt gene is wild-type.

14. The method of claim 11, wherein the fusion protein comprises a nuclease domain and expression of the Htt gene is inactivated.

15. A cell made by the method of claim 11, wherein the cell comprises the one or more polynucleotides.

16. A method of modifying an Htt gene in a cell, the method comprising, administering to the cell one or more polynucleotides encoding one or more fusion proteins according to claim 9, wherein the one or more fusion proteins comprises a nuclease domain and wherein the sequence of the Htt gene is modified.

17. A method of generating a model system for the study of Huntington'"'"'s Disease, the method comprising modifying an Htt gene according to the method of claim 16.

18. The method of claim 17, wherein the cell is an embryonic stem cell.

19. The method of claim 17, wherein the Htt gene is modified to comprise one or more mutant alleles.

20. The method of claim 19, wherein the mutant alleles comprise expanded trinucleotide repeats.