Compositions and methods for caspase-induced apoptosis

• US 20030035789A1
• Filed: 06/13/2002
• Published: 02/20/2003
• Est. Priority Date: 12/16/1999
• Status: Abandoned Application

• Current Assignee: University of Louisville Research Foundation
• Original Assignee: University of Louisville Research Foundation
• Inventors: Shirwan, Haval
• Application Number:
  US10/171,417
• Publication Number:
  US 20030035789A1
• Time in Patent Office: Days
• US Class Current: 424/85.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4747 Apoptosis related proteins
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 9/6475 Interleukin 1-beta convertase-like enzymes (3.4.22.10; 3.4.22.36; 3.4.22.63)

Abstract

This invention provides methods and compositions for caspase-induced apoptosis. Chimeric proteins are provided that target cells in which apoptosis is to be induced. The chimeric proteins of this invention activate endogenous caspase or provide caspase units that cause cell death. Methods are provided for the use of the chimeric proteins of this invention for in vivo and ex vivo therapies to treat pathological conditions.

Claims

1. A method for effecting caspase-induced apoptosis in a target cell comprising:

(a) transfecting a production cell with a vector comprising DNA encoding a protein that selectively binds to the target cell, operably linked to DNA encoding an caspase-related molecule that induces apoptosis when taken into the target cell, wherein the production cell is capable of transcribing and translating the DNA to produce chimeric proteins;

(b) recovering such chimeric proteins; and

(c) introducing an effective amount of the chimeric protein into the target cell.

2. The method of claim 1, wherein the cell is an activated lymphocyte and the caspase-related protein is Il-2, IL-4, IL-8, IL-10, IL-12, IL-15, TNF-α

, GM-CSF or TGF-β and

the apoptosis inducing molecule is one of caspase 1 to 14, or a caspase activating factor.

3. The method of claim 1, wherein the production cell is a Drosophila cell.

4. A chimeric protein comprising a protein that selectively binds to a target cell and a caspase-related apoptosis-inducing molecule.

5. The chimeric protein of claim 4, wherein the protein selectively binds to an activated lymphocyte and the apoptosis inducing molecule is IL-2, IL-4, IL-8, IL-10, IL-12, IL-15, TNF-α

, GM-CSF or TGF-β and

the apoptosis inducing molecule is one of caspase 1 to 14, or a caspase-activating factor.

6. A method of inducing allograft tolerance comprising the administration of an effective amount of the chimeric protein of claim 5.

7. A method of purging a subject of activated lymphocytes comprising contacting the subject'"'"'s lymphocytes with the chimeric protein of claim 5 ex vivo and returning the purged lymphocytes to the subject.

8. The method according to claim 7, wherein the subject has an autoimmune disease or allergies.

9. An isolated and purified nucleic acid encoding a polypeptide, wherein the polypeptide comprises a receptor-binding means and an apoptosis-inducing means.

10. The nucleic acid of claim 9, wherein the receptor-binding means comprises IL-2, Il-4, IL-8, IL-10, IL-12, IL-15, TNF-α

, GM-CSF, or TGF-β

.

11. The nucleic acid of claim 9, wherein the apoptosis-inducing means comprises any of caspases 1 to 14 or a caspase activating factor.

16. A cell comprising the nucleic acid of claim 9.

19. A method of inducing apoptosis in a cell comprising contacting the cell with the nucleic acid of claim 9.

21. A method of inducing apoptosis in a cell comprising contacting the cell with the cell of claim 16.

24. The method of claim 19, 20, 21, 22 or 23, wherein the cell is a lymphocyte or a neoplastic cell.

25. The method of claim 24, wherein the lymphocyte is an activated T lymphocyte.

26. The method of claim 25, wherein the activated T lymphocyte is an activated T lymphocyte in, on, or from a human subject.

27. The method of claim 26, wherein the subject is in need of relief from any of the group consisting of autoimmune disease, graft-versus-host disease, allograft rejection, and xenograft rejection.

28. The method of claim 26, wherein the subject is at risk of autoimmune disease, graft-versus-host disease, allograft rejection, or xenograft rejection.

29. The method of claim 24, wherein the neoplastic cell is a human neoplastic cell.

12. A polypeptide comprising a receptor-binding means and an apoptosis-inducing means.

13. The polypeptide of claim 12, wherein the receptor-binding means comprises IL-2, Il-4, IL-8, IL-10, IL-12, IL-15, TNF-α

, GM-CSF, or TGF-β

.

14. The polypeptide of claim 12, wherein the apoptosis-inducing means comprises any of caspases 1 to 14 or a caspase activating factor.

15. The polypeptide of claim 12, wherein the apoptosis-inducing means induces apoptosis in a cell after the polypeptide has been internalized into the cell.

17. A cell comprising the polypeptide of claim 12.

18. A matrix comprising the polypeptide of claim 12.

20. A method of inducing apoptosis in a cell comprising contacting the cell with the polypeptide of claim 12.

22. A method of inducing apoptosis in a cell comprising contacting the cell with the cell of claim 17.

23. A method of inducing apoptosis in a cell comprising contacting the cell with the matrix of claim 18.