TREATMENT OF CHRONIC PAIN WITH ZINC FINGER PROTEINS

• US 20090215878A1
• Filed: 02/06/2009
• Published: 08/27/2009
• Est. Priority Date: 02/08/2008
• Status: Abandoned Application

• Current Assignee: Sangamo Biosciences, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: MILLER, JEFFREY C., TAN, SIYUAN, ZHANG, STEVE H., GREGORY, PHILIP D.
• Application Number:
  US12/366,927
• Publication Number:
  US 20090215878A1
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity

Abstract

A variety of zinc finger proteins (ZFPs) and methods utilizing such proteins are provided for use in treating chronic pain. ZFPs that bind to a target site in genes that are aberrantly expressed in subjects having chronic pain are described. In addition, ZFPs that bind to a target site in genes expressed at normal levels in subjects experiencing chronic pain, modulation of whose expression results in decreased pain perception, are also provided. For example, genes that are over-expressed in the dorsal root ganglia (DRG) of pain patients (e.g., Nav1.8) can be repressed.

Claims

1. A polynucleotide encoding a protein comprising an engineered zinc finger DNA-binding domain, wherein the DNA-binding domain comprises six zinc fingers denoted F1 through F6 in order from N-terminus to C-terminus and the amino acid sequence of recognition regions F1, F3, F5, and F6 of the zinc fingers is as follows:

2. The polynucleotide of claim 1, wherein the amino acid sequence of recognition regions F2 and F4 is as follows:

3. The polynucleotide of claim 1, wherein the amino acid sequence of recognition region F2 is selected from the group consisting of YSRGLWA (SEQ ID NO:

16), WPGSLSN (SEQ ID NO;

17), WRPNLVA (SEQ ID NO;

18), APRYLWQ (SEQ ID NO;

19), LLKYLAT (SEQ ID NO;

20), SSRYLWQ (SEQ ID NO;

23), HPRYLWQ (SEQ ID NO;

24), QRRYLWA (SEQ ID NO;

26), and QKRYLWQ (SEQ ID NO;

28), and the amino acid sequence of recognition region F4 is selected from the group consisting of TNQNRIT (SEQ ID NO;

12), LKRTLMV (SEQ ID NO;

21), LLQTLSS (SEQ ID NO;

22), LHRTLTV (SEQ ID NO;

25), VRCNLTK (SEQ ID NO;

27), LRRTLHM (SEQ ID NO;

29), and LKNALR1 (SEQ ID NO;

30).

12. The polynucleotide of claim 1, further comprising a transcriptional repression domain.

13. The polynucleotide of claim 12, wherein the transcriptional repression domain is a KRAB domain.

14. The polynucleotide of claim 1, further comprising a transcriptional activation domain.

15. A polypeptide encoded by the polynucleotide of claim 1.

16. The polypeptide of claim 15, further comprising a transcriptional repression domain.

17. The polypeptide of claim 16, wherein the transcriptional repression domain is a KRAB domain.

18. The polypeptide of claim 15, further comprising a transcriptional activation domain.

4-11. -11. (canceled)

20-21. -21. (canceled)

22. A zinc finger protein comprising a zinc finger comprising an amino sequence shown in Table 1, or a nucleic acid encoding the protein.

19. The zinc finger protein of claim 22 selected from any one of the proteins provided in Table 1, or a nucleic acid encoding the protein.

23. (canceled)

24. A method for treating chronic pain in a subject, the method comprising:

introducing a nucleic acid into a subject, wherein the nucleic acid encodes a polypeptide, the polypeptide comprising;

(i) a zinc finger DNA-binding domain that is engineered to bind to a target site in Nav10.8, the zinc finger DNA-binding domain comprising six zinc fingers denoted F1 through F6 in order from N-terminus to C-terminus and wherein the amino acid sequence of recognition regions F1, F3, F5, and F6 of the zinc fingers is as follows;

25. The method of claim 24, wherein the target site in the Nav1.8 gene comprises a nucleic acid sequence of 5′

-GAAGAAgAATGAGAAGATG (SEQ ID NO;

2).

26. The method of claim 24, wherein the transcriptional repression domain is a KRAB domain.

27. The method of claim 24, wherein the amino acid sequence of recognition regions F2 and F4 is as follows:

28. The method of claim 24, wherein the amino acid sequence of recognition region F2 is selected from the group consisting of YSRGLWA (SEQ ID NO:

16), WPGSLSN (SEQ ID NO;

17) WRPNLVA (SEQ ID NO;

18), APRYLWQ (SEQ ID NO;

19), LLKYLAT (SEQ ID NO;

20) SSRYLWO (SEQ ID NO;

23), HPRYLWQ (SEQ ID NO;

24), QRRYLWA (SEQ ID NO;

26), and QKRYLWQ (SEQ ID NO;

28), and the amino acid sequence of recognition region F4 is selected from the group consisting of TNQNRIT (SEQ ID NO;

12), LKRTLMV (SEQ ID NO;

21), LLQTLSS (SEQ ID NO;

22), LHRTLTV (SEQ ID NO;

25), VRCNLTK (SEQ ID NO;

27), LRRTLHM (SEQ ID NO;

29), and LKNALR1 (SEQ ID NO;

30).

70. The method of claim 24, wherein the chronic pain comprises neuropathic pain.

29-69. -69. (canceled)

71. (canceled)