Sonogenetic flexible nanogels enabling enhanced calcium overload-induced immunogenic cell death for tumor immunotherapy

Sonogenetics represents a non-invasive and precise strategy for cancer therapy by coupling ultrasound (US) with genetic engineering. However, achieving efficient deep-tumor delivery and profound immune activation remains challenging. Herein, we developed a novel sonogenetic nanogel system (NGs@pDNA) designed to trigger a potent systemic antitumor immune response via US-induced calcium (Ca 2+ ) overload. These nanogels feature flexible mechanical properties and glutathione (GSH) responsiveness, facilitating deep tumor penetration and the targeted delivery of plasmids encoding the mechanosensitive channel MscL. Upon US irradiation, the activated MscL channels drive a massive influx of endogenous Ca 2+ (5.8 ± 0.6-fold increase), disrupting intracellular homeostasis and provoking a synergistic cascade of oxidative endoplasmic reticulum (ER) stress and mitochondrial dysfunction efficiently triggering immunogenic cell death (ICD). Further, ICD promotes dendritic cell (DC) maturation and reprograms pro-tumoral M2 macrophages into the anti-tumoral M1 phenotype. Whole-genome transcriptomic sequencing reveals the significant enrichment of pathways related to Ca 2+ homeostasis, oxidative stress-induced apoptosis, and antigen processing. In vivo studies further demonstrated this sonogenetic platform effectively eradicated primary tumors and suppressed distant metastases by activating a robust systemic immune response. Overall, this study establishes a spatiotemporally controllable therapeutic paradigm by integrating sonogenetic modulation, nanogel delivery, and Ca 2+ overload-induced ICD to overcome tumor immunosuppression and enhance systemic antitumor immunity. Highly flexible NGs@pDNA penetrates the dense tumor extracellular matrix for deep plasmid delivery. After lysosomal escape and GSH-responsive plasmid release, transfected MscL enables ultrasound-triggered Ca 2+ overload, inducing mitochondrial damage, ER stress, Caspase-3 activation, and ICD, ultimately boosting systemic immune activation and antitumor immunity. • A sonogenetic nanogel system is developed for ultrasound-triggered MscL activation and endogenous Ca 2+ overload. • Crosslinking optimization endows nanogels with adaptive deformability for deep tumor penetration and transfection. • Ca 2+ influx induced by ultrasound activates an ER stress–mitochondrial dysfunction apoptotic cascade. • Sonogenetic treatment induces immunogenic cell death and remodels the tumor immune microenvironment.