Methods and compositions for treating trinucleotide repeat disorders

• US 20110082093A1
• Filed: 07/28/2010
• Published: 04/07/2011
• Est. Priority Date: 07/28/2009
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Inventors: REBAR, EDWARD J., GREGORY, PHILIP D., ZHANG, H. STEVE
• Granted Patent: US 9,234,016 B2
• Time in Patent Office: Days
• US Class Current: 514/21.2
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• 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 methods and compositions for treating trinucleotide repeat disorders.

Claims

1. An engineered zinc finger protein that modulates expression of a gene associated with a trinucleotide repeat disorder, the zinc finger protein comprising a plurality of zinc finger domains, wherein at least one zinc finger domain comprises at least one non-naturally occurring recognition helix region.

2. The zinc finger protein of claim 1, wherein the gene is wild-type.

3. The zinc finger protein of claim 1, wherein the gene comprises one or more expanded trinucleotide repeats.

4. A multimer comprising at least two zinc finger proteins of claim 1, wherein the zinc finger proteins of the multimer preferentially bind to expanded trinucleotide tracts as compared to wild-type trinucleotide repeat tracts.

5. The zinc finger protein of claim 3, wherein the expanded trinucleotide repeat is in a Huntington'"'"'s Disease (HD) gene.

6. The zinc finger protein of claim 5, wherein the HD gene is an Htt allele.

7. The zinc finger protein of claim 3, wherein the expanded trinucleotide repeat encodes poly-glutamine.

8. The zinc finger protein of claim 3, wherein the expanded trinucleotide repeat encodes poly-serine.

9. The zinc finger protein of claim 3, wherein the zinc finger protein binds to a SNP associated with the expanded trinucleotide repeat.

10. The zinc finger protein of claim 2, wherein the zinc finger protein binds to a SNP associated with a wild-type allele.

11. A fusion protein comprising a zinc finger protein according to claim 1 and a regulatory domain, a cleavage domain or a cleavage half-domain.

12. The fusion protein of claim 11, wherein the zinc finger protein activates expression of the gene.

13. The fusion protein of claim 11, wherein the zinc finger protein represses expression of the gene.

14. A polynucleotide encoding a zinc finger protein according to claim 1.

15. A polynucleotide encoding a fusion protein according to claim 11.

16. An isolated cell comprising a zinc finger protein of claim 1.

17. An isolated cell comprising a polynucleotide according to claim 14.

18. A pharmaceutical composition comprising a zinc finger protein of claim 1.

19. A pharmaceutical composition comprising a polynucleotide according to claim 14.

20. A method of targeted modification of expanded trinucleotide repeats in the genome of a cell, the method comprising administering one or more fusion proteins according to claim 11 to a cell comprising a genome with a gene comprising expanded trinucleotide repeats, such that the activity of the fusion protein results in modification of the genomic expanded trinucleotide repeats.

21. The method of claim 20, wherein the cell is selected from the group consisting of induced pluripotent stem cells (iPSC), human embryonic stem cells (hES), mesenchymal stem cells (MSC), neuronal stem cells and combinations thereof.

22. The method of claim 20, wherein the modification comprises reducing the number of trinucleotide repeats.

23. A method of generating a model system for the study of trinucleotide repeat disorders, the method comprising modifying cells such that the genome of the cells contain trinucleotide expansion tracts of specific lengths using a fusion protein according to claim 11.

24. The method of claim 23, wherein the model system comprises a cell line.

25. The method of claim 23, wherein the model system comprises a transgenic animal.

26. A method of treating a trinucleotide disorder in a subject, the method comprising;

administering at least one zinc finger protein to the subject in need of treatment, wherein the zinc finger protein modulates expression of a gene associated with the trinucleotide disorder, and further wherein the zinc finger protein is selected from the group consisting of a zinc finger protein according to claim 1, a zinc finger protein that modulates expression of a neurotrophic factor and combinations thereof.

27. The method of claim 26, wherein the neurotrophic factor comprises BDNF or GNDF.

28. The method of claim 26, wherein the gene comprises one or more expanded trinucleotide repeats and expression of the gene is repressed.