PMID- 33896272 OWN - NLM STAT- MEDLINE DCOM- 20220222 LR - 20220830 IS - 2472-5560 (Electronic) IS - 2472-5552 (Print) IS - 2472-5552 (Linking) VI - 26 IP - 7 DP - 2021 Aug TI - Sensitivity of Oncogenic KRAS-Expressing Cells to CDK9 Inhibition. PG - 922-932 LID - 10.1177/24725552211008853 [doi] AB - Oncogenic forms of KRAS proteins are known to be drivers of pancreatic, colorectal, and lung cancers. The goal of this study is to identify chemical leads that inhibit oncogenic KRAS signaling. We first developed an isogenic panel of mouse embryonic fibroblast (MEF) cell lines that carry wild-type RAS, oncogenic KRAS, and oncogenic BRAF. We validated these cell lines by screening against a tool compound library of 1402 annotated inhibitors in an adenosine triphosphate (ATP)-based cell viability assay. Subsequently, this MEF panel was used to conduct a high-throughput phenotypic screen in a cell viability assay with a proprietary compound library. All 126 compounds that exhibited a selective activity against mutant KRAS were selected and prioritized based on their activities in secondary assays. Finally, five chemical clusters were chosen. They had specific activity against SW620 and LS513 over Colo320 colorectal cancer cell lines. In addition, they had no effects on BRAF(V600E), MEK1, extracellular signal-regulated kinase 2 (ERK2), phosphoinositide 3-kinase alpha (PI3Kalpha), AKT1, or mammalian target of rapamycin (mTOR) as tested in in vitro enzymatic activity assays. Biophysical assays demonstrated that these compounds did not bind directly to KRAS. We further identified the mechanism of action and showed that three of them have CDK9 inhibitory activity. In conclusion, we have developed and validated an isogenic MEF panel that was used successfully to identify RAS oncogenic or wild-type allele-specific vulnerabilities. Furthermore, we identified sensitivity of oncogenic KRAS-expressing cells to CDK9 inhibitors, which warrants future studies of treating KRAS-driven cancers with CDK9 inhibitors. FAU - Lai, Lick Pui AU - Lai LP AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. FAU - Brel, Viviane AU - Brel V AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Sharma, Kanika AU - Sharma K AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. FAU - Frappier, Julia AU - Frappier J AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Le-Henanf, Nadia AU - Le-Henanf N AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Vivet, Bertrand AU - Vivet B AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Muzet, Nicolas AU - Muzet N AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Schell, Emilie AU - Schell E AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Morales, Renaud AU - Morales R AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Rooney, Eamonn AU - Rooney E AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Basse, Nicolas AU - Basse N AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Yi, Ming AU - Yi M AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. FAU - Lacroix, Frederic AU - Lacroix F AD - Sanofi, Molecular Oncology, Vitry-sur-Seine, France. FAU - Holderfield, Matthew AU - Holderfield M AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. FAU - Englaro, Walter AU - Englaro W AD - Sanofi, Open Innovation Access Platform, Strasbourg, France. FAU - Marcireau, Christophe AU - Marcireau C AD - Sanofi, Molecular Oncology, Vitry-sur-Seine, France. FAU - Debussche, Laurent AU - Debussche L AD - Sanofi, Molecular Oncology, Vitry-sur-Seine, France. FAU - Nissley, Dwight V AU - Nissley DV AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. FAU - McCormick, Frank AU - McCormick F AD - National Cancer Institute (NCI) RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD, USA. AD - UCSF Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, CA, USA. LA - eng GR - 75N91019D00024/CA/NCI NIH HHS/United States GR - R35 CA197709/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20210424 PL - United States TA - SLAS Discov JT - SLAS discovery : advancing life sciences R & D JID - 101697563 RN - 0 (Protein Kinase Inhibitors) RN - EC 2.7.11.22 (Cdk9 protein, mouse) RN - EC 2.7.11.22 (Cyclin-Dependent Kinase 9) RN - EC 3.6.5.2 (Hras protein, mouse) RN - EC 3.6.5.2 (Proto-Oncogene Proteins p21(ras)) SB - IM MH - Animals MH - Cyclin-Dependent Kinase 9/*antagonists & inhibitors MH - *Drug Discovery/methods MH - *Drug Screening Assays, Antitumor/methods MH - Gene Expression Regulation, Neoplastic/*drug effects MH - High-Throughput Screening Assays MH - Mice MH - Protein Kinase Inhibitors/*pharmacology MH - Proto-Oncogene Proteins p21(ras)/*genetics/metabolism PMC - PMC8791284 MID - NIHMS1770058 OTO - NOTNLM OT - CDK9 OT - KRAS OT - isogenic cell panel OT - phenotypic high-throughput screen OT - synthetic lethality EDAT- 2021/04/27 06:00 MHDA- 2022/02/23 06:00 PMCR- 2022/08/01 CRDT- 2021/04/26 05:22 PHST- 2021/04/27 06:00 [pubmed] PHST- 2022/02/23 06:00 [medline] PHST- 2021/04/26 05:22 [entrez] PHST- 2022/08/01 00:00 [pmc-release] AID - S2472-5552(22)06728-4 [pii] AID - 10.1177/24725552211008853 [doi] PST - ppublish SO - SLAS Discov. 2021 Aug;26(7):922-932. doi: 10.1177/24725552211008853. Epub 2021 Apr 24.