An actionable axis linking NFATc2 to EZH2 controls the EMT-like program of melanoma cells

Discovery of new actionable targets and functional networks in melanomais an urgent need as only a fraction of metastatic patients achieves durable clinical benefit by targeted therapy or immunotherapy approaches. Here we show that NFATc2expression is associated with an EMT-like transcriptional program and with an invasive melanomaphenotype, as shown by analysis of melanomacell lines at the mRNA and protein levels, interrogation of the TCGA melanomadataset and characterization of melanomalesions by immunohistochemistry. Gene silencing or pharmacological inhibition of NFATc2downregulated EMT-related genes and AXL, and suppressed c-Myc, FOXM1, and EZH2. Targeting of c-Myc suppressed FOXM1and EZH2, while targeting of FOXM1suppressed EZH2. Inhibition of c-Myc, or FOXM1, or EZH2downregulated EMT-related gene expression, upregulated MITF and suppressed migratory and invasive activity of neoplastic cells. Stable silencing of NFATc2impaired melanomacell proliferation in vitro and tumor growth in vivo in SCID mice. In NFATc2+ EZH2+ melanomacell lines pharmacological co-targeting of NFATc2and EZH2exerted strong anti-proliferative and pro-apoptotic activity, irrespective of BRAF or NRAS mutations and of BRAF inhibitorresistance. These results provide preclinical evidence for a role of NFATc2in shaping the EMT-like melanomaphenotype and reveal a targetable vulnerability associated with NFATc2and EZH2expression in melanomacells belonging to different mutational subsets.