铁死亡相关的癌症治疗受到了Fe2+/Fe3+氧化还原对和过氧化氢(H2O2)生成致命的羟基自由基(•OH)的限制。虽然外源性铁或产生ROS的药物可以增强铁死亡，但利用在铁蛋白中储存的内源性铁(活性铁库，LIP)和促进ROS生成可能更安全。在此，我们开发了一种无金属和药物的纳米医学技术，用于响应性地释放LIP和在线粒体膜上生成H2O2，增加羟基自由基的产生，增强铁死亡介导的抗肿瘤效应。我们开发了一种GSH/pH双活化的氟化交联聚乙烯亚胺与双醛聚乙二醇层纳米复合物载荷MTS-KR-SOD(Mitochondria-targeting-sequence-KillerRed-Superoxide Dismutase)和CRISPR/Cas9-CA IX(Carbonic anhydrase IX)质粒(FP@MC)，用于通过内源性铁解藕和原位ROS放大增强铁死亡。两个质粒被构建用于特异性敲低碳酸酐酶 IX 和在线粒体膜上翻译 KillerRed-SuperOxide Dismutase 重组蛋白。CA IX敲低使细胞内环境酸化，导致LIP从铁蛋白中释放出来作为“信号”，启动内源性化学动力疗法。同时，MTS-KR-SOD在590 nm激光照射下生成H2O2，协助化学动力疗法，导致ROS放大，损伤线粒体和脂质过氧化物积累。联合治疗效果加重了癌症铁死亡并抑制了肿瘤生长，为增强ROS和铁离子以促进铁死亡相关的癌症治疗提供了一个新的范式。本文受版权保护，版权所有。

Ferroptosis-related cancer therapy is limited by insufficient Fe2+ /Fe3+ redox pair and hydrogen peroxide (H2 O2 ) for producing lethal hydroxyl radicals (•OH). Although exogenous iron or ROS-producing drugs can enhance ferroptosis, exploiting endogenous iron (labile iron pool, LIP) stored in ferritin and promoting ROS generation may be safer. Herein, a metal/drug-free nanomedicine was developed for responsive LIP release and H2 O2 generation on the mitochondria membranes, amplifying hydroxyl radical production to enhance ferroptosis-mediated anti-tumor effects. A GSH/pH dual activatable fluorinated and crosslinked polyethyleneimine with dialdehyde polyethylene glycol layer nanocomplex loaded with MTS-KR-SOD (Mitochondria-targeting-sequence-KillerRed-Superoxide Dismutase) and CRISPR/Cas9-CA IX (Carbonic anhydrase IX) plasmids (FP@MC) were developed for enhanced ferroptosis through endogenous iron de-hijacking and in situ ROS amplification. Two plasmids were constructed to knock down carbonic anhydrase IX and translate KillerRed-SuperOxide Dismutase recombinant protein specifically on mitochondria membranes, respectively. The CA IX knockdown acidified the intracellular environment, leading the release of LIP from ferritin as a "flare" to initiate endogenous chemodynamic therapy. Meanwhile, MTS-KR-SOD generated H2 O2 when irradiated by a 590 nm laser to assist chemodynamic therapy, leading to ROS amplification for mitochondria damage and lipid peroxide accumulation. The combined therapeutic effects aggravated cancer ferroptosis and suppressed tumor growth, providing a new paradigm for amplifying ROS and iron ions to promote ferroptosis-related cancer therapy. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.