Abstract A026: EGF/EGFR signaling in priming for resistance to BRAF/MEK inhibitors in melanoma

Abstract

Abstract Targeted therapies kill cancer cells very efficiently. However, the effectiveness of targeted therapies is limited by the eventual emergence of resistance, arising from a small subset of cells capable of surviving therapy. Our work has shown genetically homogeneous cells, in distinct transcriptomic states, are capable of surviving combination BRAF and MEK inhibition. In the presence of targeted therapies these rare cells transform into resistant cells, we therefore call these cells ‘primed’ for resistance. Our research has focused on characterizing these primed cells, however, the mechanisms of survival remain unknown. Since primed cells overexpress receptor tyrosine kinases (RTKs), which are associated with growth and survival, we tested if overexpression of the RTK EGFR could explain priming and subsequent resistance. While melanoma cells in which we artificially overexpressed EGFR had no significant increase in resistance, EGFR overexpressing cells in the presence of exogenous EGF are highly resistant to combination BRAF/MEK inhibition. While EGF is not overexpressed in primed cells, preliminary data suggests that while most cells reduce EGF expression in the presence of BRAF/MEK inhibition, primed cells maintain their baseline expression, potentially maintaining EGF/EGFR signaling in these colonies. We are therefore testing if EGF/EGFR signaling is necessary in vitro by knocking out EGF or EGFR. Furthermore, to test if EGF/EGFR signaling can create a primed state, we used flow cytometry to determine expression of the prime cell marker NT5E. In cells with confirmed EGFR overexpression we observed an increased fraction of NT5E overexpressing cells, suggesting that EGF/EGFR signaling could initiate aspects of primeness beyond simple survival. Further research will determine if EGF/EGFR signaling can initiate an overall primed transcriptional state. The heritability of any primeness as a consequence of EGF/EGFR signaling will inform us if this signaling is a potential cause of priming or a mechanism by which paracrine signaling and/or stromal protection can eventually result in independent resistance. Citation Format: Robert J. Vander Velde, Sydney M. Shaffer. EGF/EGFR signaling in priming for resistance to BRAF/MEK inhibitors in melanoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A026.