Methods and compositions for treating Huntington's Disease

• US 10,265,377 B2
• Filed: 10/14/2016
• Issued: 04/23/2019
• Est. Priority Date: 02/29/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Miller, Jeffrey C., Rebar, Edward J., Zhang, H. Steve
• Primary Examiner(s): Weidner, Adam
• Application Number:
  US15/294,062
• Publication Number:
  US 20170035839A1
• Time in Patent Office: 921 Days
• Field of Search: None
• 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 method of binding a zinc finger protein to a target site in an Htt gene in a cell, the target site comprising the sequence CAGCAG, CTGCTG, GCAGCA, GCTGCT, AGC, CAG, CTG, GCA, GCT or AAG, the method comprising providing to the cell a non-naturally occurring zinc finger protein comprising 4, 5or 6 zinc finger domains ordered F1 to F4, F1 to F5 or F1 to F6, respectively, wherein the zinc finger protein binds to the Htt gene and further wherein positions F2 or F2 and F3 of the protein are selected from the group consisting of:

(i) F2 and F3 of the zinc finger protein comprise the recognition helix regions shown in SEQ ID NO;

161, 162, 163, 164, 165, 166 or 167 to bind to the target sequence CAGCAG;

(ii) F2 and F3 of the zinc finger protein comprise the recognition helix regions shown in SEQ ID NO;

168 to bind to the target sequence CTGCTG;

(iii) F2 and F3 of the zinc finger protein comprise the recognition helix regions shown in SEQ ID NO;

169, 170, 171 or 172 to bind to the target sequence GCAGCA;

(iv) F2 and F3 of the zinc finger protein comprise the recognition helix regions shown in SEQ ID NO;

173 or 174 to bind to the target sequence GCTGCT;

(v) F2 comprises the recognition helix region shown in SEQ ID NO;

175 to bind to the target sequence AGC;

(vi) F2 comprises the recognition helix region shown in SEQ ID NO;

176 or 177 to bind to the target sequence CAG;

(vii) F2 comprises the recognition helix region shown in SEQ ID NO;

42 or 148 to bind to the target sequence CTG;

(viii) F2 comprises the recognition helix region shown in SEQ ID NO;

18 or 178to bind to the target sequence GCA; and

(ix) F2 comprises the recognition helix region shown in SEQ ID NO;

31 to bind to the target sequence GCT.

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

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

4. The method of claim 1, wherein the zinc finger protein further comprises a transcriptional activation domain or a transcriptional repression domain.

5. The method of claim 4, wherein the zinc finger protein further comprises a nuclease domain.

6. The method of claim 1, wherein the zinc finger protein is provided to the cell in polynucleotide form.

7. The method of claim 1, wherein the Htt gene comprises at least one mutant allele.

8. The method of claim 1, wherein the Htt gene is wild-type.

9. The method of claim 5, wherein expression of the Htt gene is inactivated.

10. A method of modifying an Htt gene in a cell, the method comprising, binding a zinc finger protein to the Htt according to the method of claim 1, wherein the sequence of the Htt gene is modified.

11. A method of treating Huntington'"'"'s Disease, the method comprising modifying an Htt gene in a cell according to the method of claim 10 in a subject in need thereof.

12. 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 10.

13. The method of claim 12, wherein the Htt gene is modified to comprise one or more mutant alleles.

14. The method of claim 13, wherein the mutant alleles comprise expanded trinucleotide repeats.

15. The method of claim 11, wherein the cell is an embryonic stem cell.