Engineering Single-Atom Iron Nanozymes with Radiation-Enhanced Self-Cascade Catalysis and Self-Supplied H2O2 for Radio-enzymatic Therapy

Abstract

Single-atom nanozymes (SAzymes), with individually isolated metal atom as active sites, have shown tremendous potential as enzyme-based drugs for enzymatic therapy. However, using SAzymes in tumor theranostics remains challenging because of deficient enzymatic activity and insufficient endogenous H₂O₂. We develop an external-field-enhanced catalysis by an atom-level engineered FeN₄-centered nanozyme (FeN₄-SAzyme) for radio-enzymatic therapy. This FeN₄-SAzyme exhibits peroxidase-like activity capable of catalyzing H₂O₂ into hydroxyl radicals and converting single-site Fe^II species to Fe^III for subsequent glutathione oxidase-like activity. Density functional theory calculations are used to rationalize the origin of the single-site self-cascade enzymatic activity. Importantly, using X-rays can improve the overall single-site cascade enzymatic reaction process via promoting the conversion frequency of Fe^II/Fe^III. As a H₂O₂ producer, natural glucose oxidase is further decorated onto the surface of FeN₄-SAzyme to yield the final construct GOD@FeN₄-SAzyme. The resulting GOD@FeN₄-SAzyme not only supplies in situ H₂O₂ to continuously produce highly toxic hydroxyl radicals but also induces the localized deposition of radiation dose, subsequently inducing intensive apoptosis and ferroptosis in vitro. Such a synergistic effect of radiotherapy and self-cascade enzymatic therapy allows for improved tumor growth inhibition with minimal side effects in vivo. Collectively, this work demonstrates the introduction of external fields to enhance enzyme-like performance of nanozymes without changing their properties and highlights a robust therapeutic capable of self-supplying H₂O₂ and amplifying self-cascade reactions to address the limitations of enzymatic treatment.