Abstract 101: Adaptive feedback reactivates MAPK signaling in KRAS-mutant cancers with inhibition of MEK, but not ERK
Authors: Ahronian Leanne G. , Misale Sandra , Godfrey Jason T. , Nishimura Koki , Chen Lifeng , Engelman Jeffrey A. , Corcoran Ryan B. 
Published: July 2017

Abstract
Activating mutations in the KRAS oncogene occur in about 40% of colorectal cancers (CRCs) and over 90% of pancreatic ductal adenocarcinomas (PDACs). Since development of small molecules capable of inhibiting KRAS directly has proven difficult, alternative strategies have instead focused on inhibiting downstream effector pathways, such as the MAPK pathway. However, inhibition of the MAPK pathway alone with MEK inhibitors, such as selumetinib and trametinib, produces only cytostatic effects and is insufficient to kill KRAS-mutant cancer cells.
We hypothesized that inhibition of an additional kinase during MEK inhibitor treatment could improve response. We performed a kinase-targeting shRNA screen to find kinases whose knockdown would cooperate with trametinib in KRAS-mutant CRC and PDAC cell lines. The kinases found in this screen represent potential therapeutic targets to inhibit in combination with MEK.
Interestingly, despite using a very high concentration of trametinib in the screen to enrich for hits outside of the MAPK pathway, the most highly ranked kinases in the screen were members of the MAPK pathway, including ARAF, BRAF, CRAF, and MEK1. This suggests that even at high concentration, trametinib produces suboptimal MAPK inhibition. Indeed, we found that while MEK inhibitors produce robust inhibition of MAPK signaling initially, pathway reactivation was observed by 48-96 hours despite regular replenishment of drug. This feedback reactivation was accompanied by marked increases in active CRAF and phosphorylated MEK. In fact, experimental approaches that artificially increased upstream signaling flux through the MAPK pathway led to a &gt;10-fold reduction in the ability of MEK inhibitors to inhibit the MAPK pathway.
Remarkably, despite triggering the same degree of adaptive upstream MAPK signaling as seen with MEK inhibitor, we found that ERK inhibitors were able to maintain MAPK pathway suppression. Importantly, these differences in MAPK pathway suppression amount to differences in cell viability. Over four weeks, ERK inhibitor treatment reduces the outgrowth of KRAS-mutant cell lines compared to those treated with MEK inhibitors. Additionally, as these inhibitors are not used as monotherapies, replacement of trametinib with an ERK inhibitor in therapeutically relevant combination treatments improved cell responses over four weeks.
Despite the feedback reactivation of the MAPK pathway, we find that ERK inhibitors are less sensitive to this signaling than MEK inhibitors, and can effectively maintain suppression of MAPK signaling. The findings of our screen demonstrate that MAPK pathway targeting is key to successful treatment of KRAS-mutant cancers, and that ERK inhibition provides greater opportunity for inactivating MAPK. Further exploration into the mechanisms of pathway feedback will be necessary to developing valuable clinical combinations for KRAS-mutant cancers.
Citation Format: Leanne G. Ahronian, Sandra Misale, Jason T. Godfrey, Koki Nishimura, Lifeng Chen, Jeffrey A. Engelman, Ryan B. Corcoran. Adaptive feedback reactivates MAPK signaling in KRAS-mutant cancers with inhibition of MEK, but not ERK [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 101. doi:10.1158/1538-7445.AM2017-101