Ultrasound-programmable tumor extracellular vesicles delivery system for dual immune-epigenetic therapy against...

Extracellular vesicles (EVs) are promising drug carriers for cancer therapy, but conventional engineering methods often suffer from limited efficacy and safety concerns. Here, we report a non-invasive strategy to generate tumor-derived EV using low-frequency focused ultrasound stimulation, termed US-EV. Brief ultrasound exposure markedly increased EV yield, enhanced tumor-homing capacity, and reduced nucleic acid cargo, thereby lowering potential nucleic acid-related safety risks. Proteomic analyses revealed activation of inflammatory and apoptosis-related pathways, accompanied by upregulation of adhesion molecules that promote EV biogenesis and targeting. Functionally, US-EV induced immunogenic cell death and efficiently co-delivered a sonosensitizer (Hematoporphyrin monomethyl ether, HMME) and YTH N6-methyladenosine RNA-binding protein 1 (YTHDF1)-targeting siRNA to tumors. Upon ultrasound activation, HMME remodeled the tumor immune microenvironment, while siYTHDF1 mediated gene silencing and epigenetic reprogramming, overcoming dendritic cell resistance and enhancing cytotoxic T cell infiltration. This combinational approach suppressed both primary and metastatic tumor growth and elicited durable systemic antitumor immunity in preclinical models. Our ultrasound-triggered EV platform provides a safe, efficient, and translational strategy for metastatic colorectal cancer therapy.