JHU/CCR Fellowship in Molecular Targets and Drug Discovery Technologies Project Details

Project Sponsor/Mentor:	Yves Pommier
Title:	Lab/Branch Chief
Address:	Bldg. 37, Rm. 5068 NIH, Bethesda, MD 20892-4255
Telephone:	301-496-5944
Fax:	301-402-0752
Email:	pommier@nih.gov
Sponsoring Laboratory/Branch:	Laboratory of Molecular Pharmacology

Project Title:	Chk2 inhibitors
Target(s) of Interest:	Cell cycle and apoptosis

Project Synopsis:
Identification of Chk2 inhibitors and determine their molecular and cellular mechanisms of action. There is currently no specific inhibitor known for Chk2, and we expect that such inhibitors will be useful for cancer therapy as well as for molecular and cellular studies. We have developed molecular and biochemical assays to measure Chk2 activity, and developed a high throughput assay to screen chemical libraries. We have cell lines with Chk2-deficiencies and mutations. Those cell lines can be used to study the effect of the potential inhibitors by themselves or in combination. Recent Chk2 publications from our laboratory: 1. Pommier Y, Sordet O, Rao A, Zhang H, Kohn KW. Targeting Chk2 kinase: molecular interaction maps and therapeutic rationale. Curr Pharm Des 2005;11:2855-72. 2. Pommier Y, Weinstein JN, Aladjem MI, Kohn KW. Chk2 molecular interaction map and rationale for Chk2 inhibitors. Clin Cancer Res 2006;12:2657-61. 3. Jobson AG, Cardellina JH, 2nd, Scudiero D, et al. Identification of a Bis-guanylhydrazone [4,4'-Diacetyldiphenylurea-bis(guanylhydrazone); NSC 109555] as a Novel Chemotype for Inhibition of Chk2 Kinase. Mol Pharmacol 2007;72:876-84. 4. Lountos GT, Tropea JE, Zhang D, et al. Crystal structure of checkpoint kinase 2 in complex with NSC 109555, a potent and selective inhibitor. Protein Science 2009;18:92-100.
Fellow Research Plan:
- Biochemical assays to determine Chk2 activity using autophosphorylation of histone H1 substrates. - Expression of recombinant Chk2 using bacterially expressed protein. - Site-directed mutagenesis to investigate specific amino acid residues on Chk2. - Cell culture to determine drug response/cytotoxicity. - siRNA experiments to identify cell lines where Chk2 deficiency has a profound impact on drug combination studies. - Measure of DNA damage and apoptosis in cell culture. - Do combination experiments with Chk2 lead inhibitors and Top1 inhibitors.