Methods and compositions for modulating PD1

• US 20110136895A1
• Filed: 11/17/2010
• Published: 06/09/2011
• Est. Priority Date: 09/27/2007
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Mendel, Matthew C., Reik, Andreas, Meng, Xiangdong, Urnov, Fyodor, Holmes, Michael C., Gregory, Philip D., Paschon, David
• Granted Patent: US 8,563,314 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 2319/71 containing domain for transcriptional activaation, e.g. VP16
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Disclosed herein are methods and compositions for modulating expression of a PD1 gene.

Claims

1. A zinc finger protein that binds to a target site specified in Table 2 (SEQ ID NOs:

56 to

     60) or Table 6 (SEQ ID NOs;

75-78) in a PD1 gene.

2. A fusion protein comprising a zinc finger protein according to claim 1 and a functional domain.

3. The fusion protein of claim 2, wherein the functional domain comprises a transcriptional regulatory domain.

4. The fusion protein of claim 3, wherein the transcriptional regulatory domain is an activation domain or a repression domain.

5. The fusion protein of claim 2, wherein the functional domain comprises a cleavage domain or cleavage half-domain.

6. A polynucleotide comprising a zinc finger protein according to claim 1.

7. A polynucleotide comprising a fusion protein according to claim 2.

8. A method of modulating expression of a PD1 gene in a cell, the method comprising

introducing into the cell at least one polynucleotide according to claim 7, under conditions such that expression of the PD1 gene is modulated.

9. The method of claim 8, wherein the fusion protein comprises an activation domain and PD1 expression is increased.

10. The method of claim 8, wherein the fusion protein comprises a repression domain and PD1 expression is decreased.

11. The method of claim 8, wherein the fusion protein comprises a cleavage domain and the PD1 gene is inactivated.

12. The method of claim 11, wherein inactivation occurs via non-homologous end joining (NHEJ) following cleavage.

13. The method of claim 11, further comprising introducing an exogenous sequence flanked by sequences having homology to the PD1 gene into the cell, wherein the exogenous sequence is integrated into the PD1 gene by homologous recombination.

14. The method of claim 8, wherein the cell is a eukaryotic cell.

15. The method of claim 14, wherein the cell is a primary cell.

16. The method of claim 14, wherein the primary cell is selected from the group consisting of peripheral blood mononuclear cells (PBMC), CD4+ T cells and CD8+ T cells.

17. The method of claim 14, wherein the cell is a stem cell.

18. The method of claim 17, wherein the stem cell is selected from the group consisting of embryonic stem cells, induced pluripotent stem cells, hematopoietic stem cells, neuronal stem cells and mesenchymal stem cells.

19. A method of treating a subject having a disease or disorder characterized by aberrant expression or regulation of a PD1 gene, the method comprising

modulating expression of the PD1 gene according to the method of claim 8 in the subject.

20. The method of claim 19, wherein expression of the PD1 gene is repressed.

21. The method of claim 19, wherein the fusion protein comprises a cleavage domain and the PD1 gene is inactivated.

22. The method of claim 19, wherein the disease or disorder is a cancer, a viral infection or an autoimmune disease.

23. The method of claim 19, further comprising administering additional anti-viral or anti-cancer therapies to the subject.

24. The method of claim 23, wherein the additional anti-viral or anti-cancer therapies are administered prior to, concurrently or sequentially with the fusion protein.