Abstract 378: HDAC inhibition enhances MEK antagonist therapy in uveal melanoma through combined blockade of YAP, AKT and RTK signaling

Abstract

Abstract Around 85-90% of all uveal melanomas harbor driver mutations in GNAQ or GNA11 leading to constitutive activation of numerous signaling pathways, including the MAPK pathway. MEK inhibitors have been evaluated clinically for metastatic uveal melanoma, but the responses are short-lived and the mechanisms of adaptation are poorly understood. In the current study, we performed RNA-seq and activity-based protein profiling (ABPP) to define the adaptive response of uveal melanoma cells to MEK inhibition and to design more effective combination therapy strategies. These analyses showed that MEK inhibition caused cytoskeleton remodeling driven by cortactin/Rho-GTPases with an increase in YAP activity, which in turn allowed therapeutic escape. Cortactin knockdown decreased YAP activity in response to MEK inhibition, increased cell death in vitro and was associated with tumor shrinkage in vivo. The proteomic data showed that MEK inhibition increased HDAC activity and an increase in global protein deacetylation. Co-targeting of HDACs and MEK was associated with increased apoptosis, decreased survival in 2D and 3D cell culture assays and suppression of YAP signaling. As YAP was unlikely to be the only escape pathway, we performed kinome and RTK arrays and demonstrated MEK inhibition also increases ROR1/2 and IGF-1R phosphorylation, leading to downstream PI3K/AKT signaling. At a signal transduction level, the combination of a pan-HDAC inhibitor (panobinostat) with a MEK inhibitor (trametinib) blocked all of the adaptive signaling pathways we identified, including RTKs, AKT, YAP and cortactin. In vivo xenograft studies revealed the MEK/HDAC inhibitor combination to outperform either agent alone, leading to long-term decrease of tumor growth and the suppression of adaptive PI3K/AKT, cortactin and YAP signaling. These findings identify HDAC inhibitors as a promising combination partner for MEK inhibitors in advanced uveal melanoma that may lead to improved systemic responses. Citation Format: Fernanda Faiao-Flores, Michael Emmons, Michael Durante, Biswarup Saha, Bin Fang, John Koomen, Srikumar Chellappan, Silvya Maria-Engler, Jonathan Licht, William Harbour, Keiran Smalley. HDAC inhibition enhances MEK antagonist therapy in uveal melanoma through combined blockade of YAP, AKT and RTK signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 378.