NCLX Inhibition plus Ca2+ Delivery Potentiates Electroporation and Antitumor Immunity in Pancreatic Ductal Adenocarcinoma

Irreversible electroporation (IRE) is increasingly used for locally advanced pancreatic ductal adenocarcinoma (PDAC), yet electric-field heterogeneity leaves a low-field rim where residual cells survive, and recurrence is frequent, compounded by a profoundly immunosuppressive tumor microenvironment. Here, an electroporation-synchronized adjuvant strategy is developed that integrates intratumoral Ca2+ delivery with nanoparticle-mediated inhibition of mitochondrial NCLX using CGP37157 (mPEG-PCL@CGP37157). This regimen markedly enhances cytotoxicity in PDAC cell lines and patient-derived organoids and significantly improves local control and survival in syngeneic subcutaneous and orthotopic PDAC models, without overt systemic toxicity. Reduced therapeutic efficacy in T cell-deficient hosts demonstrates the importance of adaptive immunity. Mechanistically, EP-resistant PDAC cells upregulate NCLX to buffer mitochondrial Ca2+ stress. Concurrent Ca2+ delivery and pharmacological NCLX inhibition induce sustained mitochondrial Ca2+ overload, triggering immunogenic GSDMD-dependent pyroptosis. This is accompanied by enhanced dendritic cell activation and CD8+ T-cell responses, as well as suppression of a tumor-myeloid SPP1-CD44 axis, promoting pro-inflammatory macrophage polarization. Collectively, targeting NCLX potentiates the immunogenic effects of IRE and provides a translatable strategy for eliminating residual disease in PDAC.