Abstract
Melanoma plasticity drives immune evasion and therapy resistance through dynamic cell-state transitions beyond genetic alterations. Although epigenetic remodeling is central to this process, its impact under therapeutic pressure remains unclear. We profiled longitudinal biopsies from melanoma patients treated in the phase Ib NIBIT-M4 epi-immunotherapy trial (NCT02608437, DNMT1 inhibitor plus anti-CTLA4) using single-cell multiome and spatial transcriptomics. Seven malignant meta-programs were identified, including a rare Wnt/β-catenin melanocytic state and a de-differentiated neural crest–like state enriched in non-responders. Spatial analyses showed that homotypic clustering stabilizes resistant programs, with neural crest–like cells forming compact niches. Responders displayed enrichment of antigen presentation/interferon programs and coordinated T and B cell expansion, whereas non-responders retained stable neural crest–like clusters. Epigenetic therapy reactivated transposable elements, priming innate immunity and enhancing immunogenicity. NFATC2 emerged as a master regulator of neural crest–like states and resistance; its perturbation promoted differentiation and immunogenicity. These findings define mechanisms of resistance and nominate β-catenin and NFATC2 as therapeutic vulnerabilities.