Methods of determining cellular chemosensitivity
The present invention provides methods of determining cell sensitivity to a therapeutic agent.
1. A method of determining whether a chemotherapeutic agent will induce cell death in cancer cells of a cancer cell sample, the method comprising:
contacting a cancer cell of the cancer cell sample with a BH3 domain peptide, wherein the cancer cell is a primary human cancer cell; and
detecting BH3 domain peptide-induced mitochondrial outer membrane permeabilization of said cancer cell,
wherein the method is performed in the absence of contacting the cancer cell with the chemotherapeutic agent and wherein the presence of BH3 domain peptide-induced mitochondrial outer membrane permeabilization indicates treatment comprising the chemotherapeutic agent will induce cell death in cancer cells of the cancer cell sample, wherein said treatment comprising the chemotherapeutic agent does not comprise treatment with a BH3 domain mimetic or an antagonist of antiapoptotic BCL-2 family members.
2. A method of determining whether a chemotherapeutic agent will induce cell death in cancer cells of a cancer cell sample, the method comprising:
contacting mitochondria from a cancer cell of the cancer cell sample with a BH3 domain peptide or mimetic thereof, wherein the cancer cell is a primary human cancer cell; and
detecting cytochrome c release from said mitochondria,
wherein the method is performed in the absence of contacting the cancer cell with the chemotherapeutic agent and wherein the cytochrome c release indicates treatment comprising the chemotherapeutic agent will induce cell death in cancer cells of the cancer cell sample, wherein said treatment comprising the chemotherapeutic agent does not comprise treatment with a BH3 domain mimetic or an antagonist of antiapoptotic BCL-2 family members.
3. The method of claim 1 , wherein said cancer cell is permeabilized prior to contacting with said BH3 domain peptide.
4. The method of claim 3 , further comprising contacting said permeabilized cell with a potentiometric dye.
5. The method of claim 4 , where said potentiometric dye is JC-1 or dihydrorhodamine 123.
6. The method of claim 4 , where said BH3 domain peptide-induced mitochondrial outer membrane permeabilization is measured by detecting a change in emission of said potentiometric dye.
7. The method of claim 1 , 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.
8. The method of claim 1 , wherein said BH3 domain peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14 and 15.
9. The method of claim 1 , wherein said cancer cell is a lymphoma cell, a leukemia cell, or a small cell lung cancer cell.
10. The method of claim 1 , wherein said BH3 domain peptide is derived from the BH3 domain of a NOXA polypeptide, and wherein the presence of BH3 domain peptide-induced mitochondrial outer membrane permeabilization indicates said cancer cell is an MCL-1-dependent cancer cell.
11. The method of claim 1 , wherein the BH3 domain peptide is less than or equal to 50 amino acids in length, less than or equal to 35 amino acids in length, less than or equal to 25 amino acids in length, or less than or equal to 15 amino acids in length.
12. The method of claim 8 , wherein the amino acid sequence comprises at least one of a tryptophan substitution or a tyrosine substitution at one or more amino acid positions.