Abstract PR013: Dynamic kinome reprogramming and metabolic rewiring drive adaptive resistance to RAS inhibition in pancreatic cancer

Abstract Direct RAS inhibitors are poised to transform the treatment landscape for pancreatic ductal adenocarcinoma (PDAC), where frontline therapy remains cytotoxic chemotherapy with limited clinical benefit. Despite this progress, intrinsic and acquired resistance limit the depth and duration of response. While putative genetic resistance mechanisms explain approximately 50% of cases in non-small cell lung cancer, colorectal cancer, and PDAC, the non-genetic mechanisms driving the remaining 50% remain poorly defined. We utilized transcriptomics, proteomics, and CRISPR-Cas9 genetic screens to elucidate the adaptive programs driving KRAS-independent growth. We identified the activation of distinct transcriptional drivers—MYC, YAP-TEAD, and KEAP1-NRF2—that bypass KRAS inhibition to sustain the resistant state. Comparative RNA-sequencing revealed that MYC- and TEAD-driven networks share substantial overlap with RAS-regulated networks, converging on essential cell cycle and growth genes. In contrast, the KEAP1-NRF2 network operates via a distinct axis characterized by increased dependence on glutamine metabolism. We confirmed the enrichment of these signatures in both preclinical models and patient samples exhibiting resistance to RAS inhibition. We then sought to identify therapeutic strategies to target these resistant states. To identify the upstream signaling governing the YAP/TEAD transcriptional shift, we performed comprehensive phosphoproteomics on PDAC cell lines treated with RAS inhibitors. We observed dynamic kinome reprogramming characterized by the activation of RHO GTPase effector kinases, specifically PAK, ROCK, and PKN. Mechanistically, we demonstrate that pharmacological inhibition of ROCK or PAK decreases nuclear YAP localization and restores sensitivity to RAS inhibition. To identify therapeutic strategies to target NRF2-driven resistance, we leveraged our transcriptomics to identify a metabolic shift with increased reliance on glutamine metabolism. Functionally targeting this metabolic vulnerability with the clinically tractable glutamine antagonists, CB-839/telaglenastat or DRP-104/sirpiglenastat, significantly impaired tumor growth in NRF2-activated models. Collectively, these findings delineate a hierarchy where kinome remodeling drives the transcriptional and metabolic plasticity required for drug tolerance, highlighting the RHO-ROCK-YAP axis and NRF2-regulated glutamine metabolism as actionable targets to extend the durability of next-generation RAS therapies. Citation Format: Clint A. Stalnecker, Wen-Hsuan Chang, Brandon L. Mouery, Ryan D. Mouery, Oluwadara Coker, A. Cole. Edwards, Runying Yang, Crystal L. Pace, Laura E. Herring, Whitney L. Stutts, Joshua H. Choe, Alec J. Vaughan, Timour Baslan, Ben Z. Stanger, Kwok-Kin Wong, Thales Y. Papagiannakopoulos, Andrew J. Aguirre, Joseph D. Mancias, Adrienne D. Cox, Channing J. Der. Dynamic kinome reprogramming and metabolic rewiring drive adaptive resistance to RAS inhibition in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RAS Oncogenesis and Therapeutics; 2026 Mar 5-8; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(5_Suppl_1):Abstract nr PR013.