Methods of Determining Cellular Chemosensitivity

• US 20080199890A1
• Filed: 04/02/2007
• Published: 08/21/2008
• Est. Priority Date: 03/31/2006
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Letai, Anthony
• Granted Patent: US 8,221,966 B2
• Time in Patent Office: Days
• US Class Current: 435/7.23
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C12Q 1/025 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics antimicrobial activity C12Q1/18
• CPC Class Codes: G01N 2510/00 Detection of programmed cell death, i.e. apoptosis
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/5748 involving oncogenic proteins

Abstract

The present invention provides methods of determining cell sensitivity to a therapeutic agent.

Claims

1. A method of predicting sensitivity of a cancer cell to a therapeutic agent comprising contacting said cell or cellular component thereof with a BH3 domain peptide and detecting apoptosis of said cell, wherein the presence of apoptosis indicate said cell is sensitive to a therapeutic agent.

5. The method of claim 1, 3 or 4 wherein said cell is permeabilized prior to contacting with said BH3 domain peptide or mimetic thereof.

6. The method of claim 5, further comprising connecting said permeabilized cell with a potentiometric dye.

7. The method of claim 6, where said potentiometric dye is JC-1 or dihydrorhodamine 123.

8. The method of claim 6, where said apoptois is measured by measured by detecting a change in emission of said potentiometric dye.

9. The method of any one of claims 1-4 wherein said BH3 domain peptide is derived from the BH3 domain of a BID, a BIM, a BAD, a BIK, a NOXA, a PUMA a BMF, or a HRK polypeptide

10. The method of any one of claims 1-4 wherein said BH3 domain peptide is selected from the group consisting of SEQ ID NO:

1-14 and 15.

11. The method of any one of claim 1-4, wherein said therapeutic agent is a chemotherapeutic agent, BH3 domain mimetic, or antagonist of an anti-apoptotic protein.

2. A method of predicting sensitivity of a cancer cell to a therapeutic agent comprising contacting mitochondria from said cell with a BH3 domain peptide or mimetic thereof and detecting cytochrome C release from said mitochondria, wherein the cytochrome C release indicates said cell is sensitive to a therapeutic agent.

3. A method of selecting an agent that is therapeutic for a subject comprising:

a) providing a cancer cell or cellular component thereof from said subject;

b) contacting said cancer cell with a BH3 domain peptide or mimetic thereof,

b) determining whether or not said BH3 domain peptide or mimetic induces apoptosis in said cancer cell to produce a test BH3 profile;

c) comparing said test BH3 profile with a therapeutic agent BH3 profile;

wherein a similarity of said test BH3 profile compared to said therapeutic agent BH3 profile indicates that the agent is therapeutic for said subject.

4. A method of predicting sensitivity of a cancer cell to a therapeutic agent comprising providing a BH3 profile of said cancer cell and comparing said BH3 profile to a control profile, wherein a similarity of said BH3 profile in said cancer cell compared to said control profile indicate said cancer cell is sensitive to said therapeutic agent.

12. A profile, comprising a pattern of mitochondrial sensitivity to BH3 peptides selected from the group consisting of SEQ ID NO:

1-12 and 13 taken from one or more subjects who have cancer.

13. A cell based assay system comprising permeabilized labeled cell and a BH3 peptide or mimetic thereof.

14. The assay system of claim 13, wherein said label comprises a potentiometric dye.

15. The method of claim 14, where said potentiometric dye is JC-1 or dihydrorhodamine 123.