A T‐cell Inspired Sonoporation System Enhances Low‐dose X‐ray‐mediated Pyroptosis and Radioimmunotherapy Efficacy by Restoring Gasdermin‐E Expression

Abstract Genome editing has the potential to improve the unsatisfactory therapeutic effect of antitumour immunotherapy. However, the cell plasma membrane prevents the entry of almost all free genome‐manipulation agents (e.g., DNAs, RNAs, proteins). Therefore, a system that can be spatiotemporally controlled and can instantly open the cellular membrane to allow the entry of genome‐editing agents into target cells is needed. Here, inspired by the ability of cytotoxic T cells to deliver cytotoxins to cancer cells by perforation, we established an easy‐to‐prepare and long shelf‐stable ultrasound (US)‐controlled perforation system (UPS) to enhance the delivery of free genome‐manipulating agents. Our UPS can precisely perforate the tumor cell membrane while maintaining cell viability via a controllable lipid peroxidation reaction. In vitro, transmembrane‐incapable plasmids can enter cells and perform genome editing with the assistance of the UPS, achieving an efficiency of up to 90%. In vivo, our UPS is biodegradable, nonimmunogenic, and tumor‐targeting, enabling the puncturing of tumor cells under US. With the application of UPS‐assisted genome editing, we successfully restored gasdermin E expression in 4T1 tumor‐bearing mice, which led to pyroptosis‐mediated antitumor immunotherapy via low‐dose X‐ray irradiation (5 Gy in total). This study provides new insights for designing a sonoporation system for genome editing. Moreover, our results demonstrated that restoring gasdermin expression by genome editing significantly improved the efficacy of radioimmunotherapy. This article is protected by copyright. All rights reserved