Thermo‐Responsive Catalytic Hydrogel Enables Oncolytic Enzyme Therapy Through Cascading Plasma Membrane Rupture and Oxidative Stress Amplification

Abstract Active initiation of plasma membrane rupture represents a promising strategy to disrupt cellular homeostasis and induce cancer cell death. This study proposes phospholipase A1 (PLA1) as a molecular “cell‐puncher” to hydrolyze phospholipids, compromising cancer cell membrane integrity and enabling uncontrolled molecular flux. PLA1 synergistically enhances the tumoradical efficacy of cholesterol oxidases (CODs) and lipoxygenases (LOXs) by liberating their respective substrates, cholesterol, and polyunsaturated fatty acids. When localized within tumors using a thermoresponsive chitosan/β‐glycerol‐phosphate hydrogel, PLA1‐COD or PLA‐LOX enzymatic pairs achieved effective treatment of CT26 murine tumors through cascading plasma membrane rupture and oxidative stress amplification. Furthermore, the hydrogel‐embedded enzyme system functioned as an injectable embolic agent, suppressing orthotopic N1S1 hepatoma in rats via transcatheter arterial enzyme embolization. This work demonstrated an enzyme‐based oncolytic strategy that targets membrane integrity and oxidative stress pathways, showing significant potential for clinical translation in solid tumor management.