METHODS OF IDENTIFYING A CANDIDATE COMPOUND
The invention relates to methods identifying compounds for the treatment of cell proliferation-related disorders, e.g., proliferative disorders such as cancer.
1 . A method of identifying a candidate compound that selectively interferes with proliferation or viability of a first IDH (isocitrate dehydrogenase)-mutant cell that has elevated levels of 2HG (2-hydroxyglutarate) comprising contacting a candidate compound with a first IDH-mutant cell that has elevated levels of 2HG, and if proliferation or viability of the first IDH-mutant cell that has elevated levels 2HG is decreased as compared to a control cell that does not have elevated levels of 2HG, then identifying the candidate compound as a compound that interferes with proliferation or viability of the first IDH-mutant cell.
2 . The method of claim 1 , wherein the first IDH-mutant cell carries a mutation in the IDH1 gene.
3 . The method of claim 1 , wherein the first IDH-mutant cell carries a mutation in the IDH2 gene.
4 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH1 R132X mutation.
5 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH2 R172X mutation.
6 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH1 R132H mutation.
7 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH2 R172K mutation.
8 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH1 R132C mutation.
9 . The method of claim 1 , wherein the first IDH-mutant cell carries an IDH2 R140Q mutation.
10 . The method of claim 1 , wherein the first IDH-mutant cell carries any one of the mutations listed in Table 1.
11 . The method of claim 1 , wherein the first IDH-mutant cell is from a cancer cell line.
12 . The method of claim 1 , wherein the first IDH-mutant cell is from a glioma cell line or a fibrosarcoma cell line.
13 . The method of claim 1 , wherein the first IDH-mutant cell is from a U87 glioma cell line.
14 . The method of claim 1 , wherein the first IDH-mutant cell is from a U87 glioma cell line engineered to express IDH1 R132H .
15 . The method of claim 1 , wherein the first IDH-mutant cell is from a fibrosarcoma /HT1080 cell line that carries an IDH1 R132C mutant gene.
16 . The method of claim 14 , wherein the control cell is a parental U87 cell.
17 . The method of claim 15 , wherein the control cell is a fibrosarcoma HT1080 cell engineered to expressed a microRNA, siRNA, or antisense RNA that inhibits expression of the IDH1 R132C mutant gene.
18 . The method of claim 1 , further comprising testing the candidate compound for tumor suppressor activity in an animal model.
19 . The method of claim 1 , further comprising administering the compound to a human who has a cancer, in an amount sufficient to treat the cancer.
20 . The method of claim 1 , wherein the cell proliferation is assayed by luminescence.
21 . The method of claim 1 , wherein the candidate compound is obtained from a library.
22 . The method of claim 1 , further comprising testing the selected candidate compound for an ability to selectively interfere with proliferation or viability of a second IDH-mutant cell comprising contacting the selected candidate compound with a second IDH-mutant cell that has elevated levels of 2HG, and if proliferation or viability of the second IDH-mutant cell is decreased as compared to a second control cell that does not have elevated levels of 2HG, then identifying the candidate compound again as a compound that selectively interferes with proliferation or viability of the first IDH-mutant cell.
23 . The method of claim 22 , wherein the first IDH-mutant cell is from a U87 cell line that carries an IDH1 R132H mutation, and the second IDH-mutant cell line is an HT1080 cell line.
24 . A method of identifying a compound that specifically interferes with an IDH (isocitrate dehydrogenase)-mutant enzyme that causes elevated levels of 2HG (2-hydroxyglutarate) comprising contacting a candidate compound with an IDH-mutant cell that has elevated levels of 2HG, and if 2HG levels are decreased as compared to a control cell that does not have elevated levels of 2HG, then identifying the candidate compound as a compound that specifically interferes with the IDH-mutant enzyme.
25 . The method of claim 24 , wherein 2-HG production is assayed by an enzymatic fluorescence assay.
26 . The method of claim 24 , further comprising testing the selected candidate compound for an ability to selectively interfere with proliferation or viability of the IDH-mutant cell, comprising contacting the selected candidate compound with the IDH-mutant cell, and if proliferation or viability of the IDH-mutant cell is decreased as compared to the control cell, then identifying the candidate compound as a compound that selectively interferes with proliferation of the IDH-mutant cell.