Method of modulating apoptosis and compositions thereof

• US 20050085420A1
• Filed: 07/22/2004
• Published: 04/21/2005
• Est. Priority Date: 07/22/2003
• Status: Abandoned Application

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Cheng, Emily, Korsmeyer, Stanley
• Application Number:
  US10/897,485
• Publication Number:
  US 20050085420A1
• Time in Patent Office: Days
• US Class Current: 514/1.400
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005

Abstract

Disclosed herein are methods for modulating apoptosis. Methods are provided for inhibiting or preventing apoptosis by promoting formation of a complex of BAK and Voltage-Dependent Anion Channel 2 (VDAC2). The invention also provides methods of promoting or inducing apoptosis by disrupting or inhibiting formation of a VDAC2/BAK complex. Also disclosed are methods of screening for compounds that promote or disrupt the VDAC2/BAK complex.

Claims

1. A method for preventing apoptosis in a cell, said method comprising contacting said cell with a wild-type VDAC2 protein, aVDAC2 mimetic or a VDAC2/BAK stabilizer compound, wherein the wild-type VDAC2 protein, VDAC2 mimetic or VDAC2/BAK stabilizer compound modulates interaction between a VDAC2 protein and a BAK protein to form a VDAC2/BAK complex, thereby preventing apoptosis.

2. The method of claim 1, wherein the cell is contacted with a VDAC2 mimetic that comprises an anti-apoptotic mutated VDAC2 peptide that binds the BAK protein.

3. A method for preventing or alleviating a symptom of an apoptosis-associated disorder in a subject, said method comprising administering a wild-type VDAC2 protein, a VDAC2 mimetic or a VDAC2/BAK stabilizer compound, wherein the wild-type VDAC2 protein, VDAC2 mimetic or VDAC2/BAK stabilizer compound modulates interaction between a VDAC2 protein and a BAK protein to form a VDAC2/BAK complex.

4. The method of claim 3, wherein the cell is contacted with a VDAC2 mimetic that comprises an anti-apoptotic mutated VDAC2 peptide that binds the BAK protein.

5. The method of claim 3, wherein the apoptosis-associated disorder is selected from the group consisting of stroke, myocardial infarction, hypertension, septic shock, organ transplantation and a neurodegenerative disorder.

6. The method of claim 5, wherein the neurodegenerative disorder is selected from the group consisting of Parkinson'"'"'s disease, amytrophic lateral sclerosis (ALS), Alzheimer'"'"'s disease, Huntington'"'"'s disease and immune deficiency.

7. A method for promoting apoptosis in a cell, said method comprising contacting said cell with a VDAC2/BAK inhibitor compound, wherein the VDAC2/BAK inhibitor compound prevents a VDAC2 protein and a BAK protein from interacting and forming a stable VDAC2/BAK complex, thereby promoting apoptosis.

8. The method of claim 7, wherein the VDAC2/BAK inhibitor compound is selected from the group consisting of a BID protein, a BIM protein, a BAD protein, a NOXA protein, a BH3 domain peptide, a BID mutein, a BIM mutein, a BAD mutein, a NOXA protein, and a BH3 domain mutein.

9. The method of claim 7, wherein the BH3 domain peptide is a BH3 only peptide.

10. The method of claim 9, wherein the BH3 only peptide is selected from the group consisting of tBID, BIM-EL and PUMA.

11. The method of claim 7, wherein the VDAC2/BAK inhibitor compound is selected from the group consisting of an anti-VDAC2 antibody, a pro-apoptotic VDAC2 mutein polypeptide and a pro-apoptotic BAK mutein polypeptide.

12. The method of claim 11, wherein the pro-apoptotic VDAC2 mutein polypeptide interacts with endogenous BAK but does not inhibit homo-oligomerization of BAK.

13. The method of claim 11, wherein the pro-apoptotic BAK mutein polypeptide interacts with endogenous VDAC2, thereby preventing endogenous VDAC2 from binding to endogenous BAK.

14. A method of preventing or alleviating a symptom of a cell-proliferative disorder, the method comprising administering a VDAC2/BAK inhibitor compound, wherein said VDAC2/BAK inhibitor compound modulates interaction between a VDAC2 protein and a BAK protein to prevent formation of a VDAC2/BAK complex, thereby promoting apoptosis in a sufficient amount to alleviate the symptom.

15. The method of claim 14, wherein the BH3 domain peptide is a BH3 only peptide.

16. The method of claim 15, wherein the BH3 only peptide is selected from the group consisting of tBID, BIM-EL and PUMA.

17. The method of claim 14, wherein the VDAC2/BAK inhibitor compound is selected from the group consisting of an anti-VDAC2 antibody, a pro-apoptotic VDAC2 mutein polypeptide and a pro-apoptotic BAK mutein polypeptide.

18. The method of claim 17, wherein the pro-apoptotic VDAC2 mutein polypeptide interacts with endogenous BAK but does not inhibit homo-oligomerization of BAK.

19. The method of claim 17, wherein the pro-apoptotic BAK mutein polypeptide interacts with endogenous VDAC2, thereby preventing endogenous VDAC2 from binding to endogenous BAK.

20. The method of claim 14, wherein the cell-proliferative disorder is selected from the group consisting of cancer, non-malignant neoplasias, DNA viruses, lymphoproliferative conditions, arthritis, inflammation and autoimmune disorders.

21. The method of claim 20, wherein the DNA virus is selected from Epstein Barr virus, African swine fever virus and adenovirus.

22. A method for screening for a VDAC2 mimetic or a VDAC2/BAK stabilizer compound, comprising the steps of:

(a) contacting a cell population with a candidate compound, wherein said cell population comprises at least one cell having a VDAC2 protein and a BAK protein; and

(b) determining the level of apoptosis of said cell population;

wherein a decrease in said level of apoptosis in the presence of the candidate compound compared to a normal control level of apoptosis in the absence of the said candidate compound indicates said candidate compound is a VDAC2 mimetic or a VDAC2/BAK stabilizer compound.

23. The method of claim 22, wherein said cell is contacted in vivo or in vitro.

24. A method for screening for a VDAC2/BAK inhibitor compound, comprising the steps of:

(a) contacting a cell population with a candidate compound, wherein said cell population comprises at least one cell having a VDAC2 protein and a BAK protein; and

(b) determining the level of apoptosis said cell population;

wherein an increase of said level of apoptosis in the presence of said candidate compound compared to a normal control level of apoptosis in the absence of said compound indicates that said candidate compound is a VDAC2/BAK inhibitor compound.

25. The method of claim 24, wherein said cell is contacted in vivo or in vitro.

26. A method of screening for an anti-apoptotic compound comprising the steps of:

(a) contacting a cell population with a candidate compound, wherein said cell population comprises at least one cell having a VDAC2 protein and a BAK protein; and

(b) determining the level of apoptosis of said cell population;

wherein a decrease in said level of apoptosis in the presence of the candidate compound compared to a normal control level of apoptosis in the absence of the said candidate compound indicates said candidate compound is an anti-apoptotic compound, further wherein the anti-apoptotic compound prevents the release of the BAK protein from a VDAC2/BAK complex, thereby inhibiting apoptosis.

27. The method of claim 26, wherein the anti-apoptotic compound is a BH3 only mutein or a small molecule compound.