Abstract 3769: Combined TRKA and MEK1/2 Inhibition Forestalls the Onset of Acquired Resistance in a New Preclinical Model of NTRK1+ Pancreatic Cancer

Abstract

Abstract NTRK1 gene fusions encode oncoprotein (TRKA) kinases that are actionable drivers of a number of human malignancies including a subset of pancreatic cancers. Here, we describe a new preclinical model of pancreatic cancer in which a TPR-NTRK1 fusion gene is expressed in conditionally immortalized mouse pancreatic ductal epithelial (IMPE) cells. Indeed, expression of TPR-NTRK1 in IMPE cells is sufficient to transform these cells without any additional engineered genetic alterations. Other oncogenes tested in this model include KRAS and EGFR, but neither were unable to replicate this one-hit model of transformation. This striking result may be explained by the differential levels of MAPK pathway activation produced by different oncogenes, and a “goldilocks zone” required to drive pancreatic cellular transformation. These data suggest NTRK1 is a unique and powerful driver of transformation in pancreatic epithelial cells. NTRK1-driven IMPE cells form fast growing tumors in immunocompromised mice both subcutaneously, and orthotopically in the mouse pancreas. Importantly, TPR-NTRK1-driven IMPE cell derived tumors are exquisitely sensitive to targeted inhibition of TRKA kinase activity. However, as also observed in pancreatic cancer patients treated with entrectinib, this therapeutic response is transient and tumors eventually develop resistance to single-agent TRKA inhibition. This model revealed that short drug treatments (2 hours) are sufficient to block autophosphorylation of the oncoprotein target, as well as blockade of the downstream MAPK signaling pathway. Critically, we observed that long-term drug treatments (24 hours) result in downstream MAPK pathway reactivation despite maintained drug activity against the target, TRKA. Consistent with this, the targeted inhibition of TRKA, which transiently blocks MAPK signaling results in a subsequent elevation of BIM protein expression, a pro-apoptotic member of the BCL2 family. Indeed, CRISPR/CAS9-mediated abrogation of BIM expression significantly diminishes the response of TPR-NTRK1-driven IMPE tumors to inhibition of TRKA kinase activity. In an effort to delay acquired resistance to single-agent TRKA inhibitors, we reasoned that combined vertical inhibition of both TRKA and MAPK signaling with the combination of an TRKA plus a MEK1/2 inhibitor might increase the durability of response in this model and counteract the observed pathway reactivation. To that end, we treated mice bearing TPR-NTRK-driven IMPE tumors with either a TRKA inhibitor, a MEK1/2 inhibitor or the combination of both therapeutic agents. Mice treated with the entrectinib/cobimetinib combination revealed the durability of the tumor regression was significantly increased compared to single-agent entrectinib. Importantly, no toxicity or adverse side effects were observed in mice receiving the combination. Additionally, the combined therapies also induced more stable expression of BIM than the single-agent, by delaying degradation of BIM by the proteasome following BIM phosphorylation. Collectively, these data provide a compelling rationale for the clinical deployment of the targeted combination of TRKA inhibition plus MEK1/2 inhibition first-line in patients whose cancers are driven by NTRK1 gene fusions. Citation Format: Aria Vaishnavi, Michael T. Scherzer, Conan Kinsey, Ignacio Garrido-Laguna, Martin McMahon. Combined TRKA and MEK1/2 Inhibition Forestalls the Onset of Acquired Resistance in a New Preclinical Model of NTRK1+ Pancreatic Cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3769.