A Synergetic Hybrid Multienzyme: Remodeling Tumor Microenvironment to Simultaneously Activate NK Cells and T Cells for Enhanced Catalytic Immunotherapy of Hepatocellular Carcinoma

ABSTRACT Hybrid multienzymes integrating natural enzymes and nanozymes are promising for reactive oxygen species (ROS)‐mediated tumor catalytic immunotherapy. However, traditional hybrid multienzymes exhibit weak catalytic synergy with insufficient ROS production. These limitations are particularly pronounced in hepatocellular carcinoma (HCC) featuring sophisticated antioxidant systems and an immunosuppressive tumor microenvironment, factors that compromise catalytic therapy and induce the dysfunction of NK and T cells. Herein, a synergetic CuSe@GOD hybrid multienzyme is constructed via in situ growth of peroxidase‐like CuSe nanozymes within the cavities of glucose oxidase (GOD). The confined intracavity catalytic space boosts CuSe nanozyme catalytic activity and enables strong catalytic synergy with GOD. In HCC cells, GOD‐produced H 2 O 2 is rapidly transformed into ROS by confined CuSe nanozymes. Meanwhile, Cu 2 + reduction‐triggered glutathione depletion and selenium species‐mediated thioredoxin reductase inactivation disrupts dual antioxidant systems to amplify ROS levels, while intracellular copper overload induces cuproptosis. Notably, ROS‐induced oxidative stress upregulates NKG2DL in HCC cells to restore NK cell function and facilitates mitochondrial DNA release during cuproptosis for cGAS‐STING pathway activation and subsequent T cell recruitment. Overall, this study establishes a synergetic hybrid multienzyme to simultaneously activate NK and T cells, offering a promising direction for HCC catalytic immunotherapy.