• Home
  • Search Results
  • Application of a MYC degradation screen identifies sensitivity to CDK9 inhibitors in KRAS-mutant pancreatic cancer.

Application of a MYC degradation screen identifies sensitivity to CDK9 inhibitors in KRAS-mutant pancreatic cancer.

Science signaling (2019-07-18)
Devon R Blake, Angelina V Vaseva, Richard G Hodge, McKenzie P Kline, Thomas S K Gilbert, Vikas Tyagi, Daowei Huang, Gabrielle C Whiten, Jacob E Larson, Xiaodong Wang, Kenneth H Pearce, Laura E Herring, Lee M Graves, Stephen V Frye, Michael J Emanuele, Adrienne D Cox, Channing J Der
ABSTRACT

Stabilization of the MYC oncoprotein by KRAS signaling critically promotes the growth of pancreatic ductal adenocarcinoma (PDAC). Thus, understanding how MYC protein stability is regulated may lead to effective therapies. Here, we used a previously developed, flow cytometry-based assay that screened a library of >800 protein kinase inhibitors and identified compounds that promoted either the stability or degradation of MYC in a KRAS-mutant PDAC cell line. We validated compounds that stabilized or destabilized MYC and then focused on one compound, UNC10112785, that induced the substantial loss of MYC protein in both two-dimensional (2D) and 3D cell cultures. We determined that this compound is a potent CDK9 inhibitor with a previously uncharacterized scaffold, caused MYC loss through both transcriptional and posttranslational mechanisms, and suppresses PDAC anchorage-dependent and anchorage-independent growth. We discovered that CDK9 enhanced MYC protein stability through a previously unknown, KRAS-independent mechanism involving direct phosphorylation of MYC at Ser62 Our study thus not only identifies a potential therapeutic target for patients with KRAS-mutant PDAC but also presents the application of a screening strategy that can be more broadly adapted to identify regulators of protein stability.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cycloheximide, from microbial, ≥94% (TLC)
Sigma-Aldrich
MG-132, Ready Made Solution, ≥90% (HPLC)
Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
Monoclonal Anti-KRAS antibody produced in mouse, clone 3B10-2F2, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
Imatinib
Sigma-Aldrich
AZ191, ≥98% (HPLC)