Gasdermin E-mediated lysosome-pore formation curbs pancreatic ductal adenocarcinoma via IFN-γ/IFN-β/TGF-β cocktail mRNA-LNP.

Despite the pleiotropic capacities of cytokines in modulating cell behaviors, their therapeutic application in cancer remains challenging. Here, we show that the IFN-γ/IFN-β/TGF-β cocktail integrates these three signals with a cytosolic pore-forming protein, gasdermin E (GSDME), and synergistically drives its delivery into the lysosomes of pancreatic adenocarcinoma (PDAC) tumor-repopulating cells (TRCs), where GSDME is cleaved to mediate lysosomal pore formation. Mechanistically, IFN-γ signaling phosphorylates GSDME, enabling phosphorylated GSDME (p-GSDME) to bind the Golgi transmembrane protein TMED10 and subsequently traffic to lysosomes, where cathepsin D cleaves it into active N-GSDME, which induces lysosomal decomposition in TRCs. In parallel, IFN-β activates STAT1/STAT3 to upregulate cathepsin D expression, whereas TGF-β enhances GSDME phosphorylation by downregulating PPP1R3G, a regulatory subunit of protein phosphatase 1. Using lipid-hybrid nanoparticle-delivered mRNA technology, the tri-cytokine cocktail demonstrated therapeutic efficacy against orthotopic PDAC in mice and PDX models, highlighting its translational potential for PDAC patients.