Abstract 5237: Systems level modeling following BRAF inhibition reveals an essential role for Ephrin A2 receptor in therapeutic escape.

Abstract

Abstract Although initially striking, the therapeutic efficacy of BRAF inhibition is eventually undermined by adaptive cellular signaling that leads to progressive disease. To date, several mechanisms of BRAF inhibitor resistance have been reported focusing primarily on single alterations. To address how global rewiring of melanoma signaling promotes resistance, we utilized mass spectrometry based phosphoproteomic and network analyses to identify signaling changes in tyrosine, threonine and serine that occur prior to and following acquired BRAF inhibitor resistance. This comprehensive bioinformatics approach uncovered a “resistance interactome” consisting of ∼150 nodes that was marked by significant alterations and the emergence of signaling nodes such as ITGB1, EphA2, EphB4, FAK1, STAT3 and PXN. GeneGo pathway enrichment analysis revealed enhanced cytoskeletal rearrangement, adhesion, integrin signaling, cell migration and extracellular matrix remodeling. Consistent with our bioinformatic predictions, melanoma cell lines with acquired vemurafenib resistance were considerably more invasive than their drug naïve counterparts and showed increased trans-endothelial cell migration. Confirmatory Western blot studies showed increased expression and phosphorylation of EphA2 receptor at Ser897, FAK1 at Ser397 and paxillin at Y118. Knockdown of EphA2 in the resistant cultures inhibited ERK signaling and completely prevented cell invasion and trans-endothelial cell migration. Functionally, EphA2 expression appeared to be regulated by an epigenetic mechanism involving promoter methylation and HDAC1. The clinical relevance of these findings was demonstrated in matched pairs of biopsies from patients receiving either BRAF or BRAF+MEK inhibitors, with significant Ephrin A2 receptor expression being observed in a subset of those on therapy. In summary, chronic MAPK pathway inhibition leads to major remodeling of the melanoma signaling network with EphA2 emerging as a key driver of the resistance phenotype. Citation Format: Kim H. T. Paraiso, Bin Fang, Dale Han, Inna Fedorenko, Jobin John, John Koomen, Jennifer Wargo, Keith Flaherty, Friedegund Meier, Kerian S. M. Smalley. Systems level modeling following BRAF inhibition reveals an essential role for Ephrin A2 receptor in therapeutic escape. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5237. doi:10.1158/1538-7445.AM2013-5237