Spatial Isolation of Single Copper(I) Sites for Cascade Enzyme-Like Catalysis and Simultaneous Ferroptosis/Cuproptosis Boosted Immunotherapy

Abstract

Nanozyme-based immunogenic cell death (ICD) inducers that effectively induce a strong immune response via enzyme-like process have attracted great attention, but how to ensure controllable active sites and maximize site utilization remains a problem. Here, we report a structurally well-defined and highly functional single-site copper(I) nanomodulators termed CuNTD, constructed by precisely anchoring atomically dispersed self-assembly S-Cu(I)-S sites onto a two-dimensional Ti₃C₂ surface. Leveraging Cu⁺ with a higher catalytic efficiency than Cu²⁺, CuNTD generates reactive oxygen species (ROS) storms through photothermal-enhanced cascade catalysis, further inducing mitochondrial dysfunction, ferroptosis and cuproptosis. Multifunctional CuNTD triggers strong ICD through cascade-regulatory pathways of photothermal-amplified ROS storms, cuproptosis and ferroptosis, effectively promoting dendritic cell maturation while reducing monotherapies side effects and resistance. In vivo, CuNTD combined with FDA-approved immunoadjuvants significantly prolong the survival of mice. With its demonstrated biosafety and high efficiency as an ICD inducer, this study provides a promising framework for advancing augmented tumor immunotherapy with significant clinical potential.

Keywords

cuproptosis / ferroptosis / immunogenic cell death / photothermal-amplified ROS storms / single-site nanozyme

References

[1] G. Bianchini, C. De Angelis, L. Licata, and L. Gianni, “Treatment Landscape of Triple-Negative Breast Cancer — Expanded Options, Evolving Needs,” Nature Reviews ClinicalOncology 19 (2022): 91.

[2] a) M. Meng, J. Wu, Y. Feng, et al., “A Comprehensive Strategy Based on High Clinical Translational Nanosystem for Programmable Immunotherapy of Triple Negative Breast Cancer,” Advanced Materials 36 (2024): 2314309. b) J. Galon and D. Bruni, “Approaches to Treat Immune Hot, Altered and Cold Tumours With Combination Immunotherapies,” Nature Reviews Drug Discovery 18 (2019): 197.

[3] a) Y. Liu, R. Niu, R. Deng, S. Song, Y. Wang, and H. Zhang, “Multi-Enzyme Co-Expressed Dual-Atom Nanozymes Induce Cascade Immunogenic Ferroptosis via Activating Interferon-γ and Targeting Arachidonic Acid Metabolism,” Journal of the American Chemical Society 145 (2023): 8965. b) Y. Liu, R. Niu, H. Zhao, et al., “Single-Site Nanozymes With a Highly Conjugated Coordination Structure for Antitumor Immunotherapy via Cuproptosis and Cascade-Enhanced T Lymphocyte Activity,” Journal of the American Chemical Society 146 (2024): 3675.

[4] Z. Zhang, Z. Pan, Q. Li, Q. Huang, L. Shi, and Y. Liu, “Rational Design of ICD-inducing Nanoparticles for Cancer Immunotherapy,” Science Advances 10 (2024): eadk0716.

[5] B. Geng, J. Hu, X. He, et al., “Single Atom Catalysts Remodel Tumor Microenvironment for Augmented Sonodynamic Immunotherapy,” Advanced Materials 36 (2024): 2313670.

[6] X. Li, J. Gao, C. Wu, et al., “Precise Modulation and Use of Reactive Oxygen Species for Immunotherapy,” Science Advances 10 (2024): eadl0479.

[7] a) J. Zhou, Q. Yu, J. Song, et al., “Photothermally Triggered Copper Payload Release for Cuproptosis-Promoted Cancer Synergistic Therapy,” Angewandte Chemie, International Edition 62 (2023): e202213922. b) C. Hao, L. Huang, H. Zhang, et al., “Chiral CuxOS@Fe-MOFs for Enhanced Cancer Therapy,” Advanced Functional Materials 34, no. 10 (2023): 2312795.

[8] a) D. Wang, H. Wu, C. Wang, et al., “Self-Assembled Single-Site Nanozyme for Tumor-Specific Amplified Cascade Enzymatic Therapy,” Angewandte Chemie, International Edition 60 (2021): 3001. b) Y. Li, Y. Dong, X. Zhou, and K. Fan, “Nanotechnology Connecting Copper Metabolism and Tumor Therapy,” MedComm 2, no. 2 (2023): e36.

[9] a) S. Zhong, C. Xiong, Y. Zhao, et al., “Self-Driven Electricity Modulates D-Band Electrons of Copper Single-Atom Nanozyme for Boosting Cancer Therapy,” Advanced Functional Materials 33 (2023): 2305625. b) H. Zhu, X. Yin, Y. Zhou, S. Xu, T. D. James, and L. Wang, “Nanoplatforms With Synergistic Redox Cycles and Rich Defects for Activatable Image-guided Tumor-Specific Therapy,” Chemistry 8 (2022): 2498.

[10] a) J. Xia, C. Hu, Y. Ji, et al., “Copper-Loaded Nanoheterojunction Enables Superb Orthotopic Osteosarcoma Therapy via Oxidative Stress and Cell Cuproptosis,” ACS Nano 17 (2023): 21134. b) G. Hou, W. Xu, M. Guo, et al., “Full-Active Cu2O/Drug Core/Shell Nanoparticles Based on “Grafting From” Drug Coordination Polymerization Combined With PD-1 Blockade for Efficient Cancer Therapy,” Chemical Engineering Journal 441 (2022): 135993. c) X. Wang, Q. Chen, Y. Zhu, et al., “Destroying Pathogen-Tumor Symbionts Synergizing With Catalytic Therapy of Colorectal Cancer by Biomimetic Protein-Supported Single-Atom Nanozyme,” Signal Transduction and Targeted Therapy 8 (2023): 277.

[11] a) P. Tsvetkov, S. Coy, B. Petrova, et al., “Copper Induces Cell Death by Targeting Lipoylated TCA Cycle Proteins,” Science 375 (2022): 1254. b) W. Xu, J. Qian, G. Hou, et al., “A Hollow Amorphous Bimetal Organic Framework for Synergistic Cuproptosis/Ferroptosis/Apoptosis Anticancer Therapy via Disrupting Intracellular Redox Homeostasis and Copper/Iron Metabolisms,” Advanced Functional Materials 32 (2022): 2205013. c) Y. Xu, S. Y. Liu, L. Zeng, et al., “An Enzyme-Engineered Nonporous Copper(I) Coordination Polymer Nanoplatform for Cuproptosis-Based Synergistic Cancer Therapy,” Advanced Materials 34 (2022): e2204733. d) L. Sun, Y. Zhang, B. Yang, et al., “Lactylation of METTL16 Promotes Cuproptosis via m6A-modification on FDX1 mRNA in Gastric Cancer,” Nature Communications 14 (2023): 6523.

[12] Z. Li, C. Wang, C. Dai, et al., “Engineering Dual Catalytic Nanomedicine for Autophagy-Augmented and Ferroptosis-Involved Cancer Nanotherapy,” Biomaterials 287 (2022): 121668.

[13] K. Chen, A. Zhou, X. Zhou, J. He, Y. Xu, and X. Ning, “Cellular Trojan Horse Initiates Bimetallic Fe-Cu MOF-mediated Synergistic Cuproptosis and Ferroptosis Against Malignancies,” Science Advances 10 (2024): eadk3201.

[14] H. Ding, F. Ren, P. Liu, et al., “Cu²⁺-Anchored Carbon Nano-Photocatalysts for Visible Water Splitting to Boost Hydrogen Cuproptosis,” Angewandte Chemie, International Edition 62 (2023): e202311549.

[15] a) D. Wang, H. Wu, S. Z. F. Phua, et al., “Self-Assembled Single-Atom Nanozyme for Enhanced Photodynamic Therapy Treatment of Tumor,” Nature Communications 11 (2020): 357. b) J. Zhou, D. Xu, G. Tian, et al., “Coordination-Driven Self-Assembly Strategy-Activated Cu Single-Atom Nanozymes for Catalytic Tumor-Specific Therapy,” Journal of the American Chemical Society 145 (2023): 4279–4293.

[16] a) N. Gong, X. Ma, X. Ye, et al., “Carbon-Dot-Supported Atomically Dispersed Gold as a Mitochondrial Oxidative Stress Amplifier for Cancer Treatment,” Nature Nanotechnology 14 (2019): 379. b) Y. Xing, J. Xiu, M. Zhou, et al., “Copper Single-Atom Jellyfish-Like Nanomotors for Enhanced Tumor Penetration and Nanocatalytic Therapy,” ACS Nano 17 (2023): 6789. c) G. Liao, L. Zhang, C. Li, S.-Y. Liu, B. Fang, and H. Yang, “Emerging Carbon-Supported Single-Atom Catalysts for Biomedical Applications,” Matter 5, no. 10 (2022): 3341–3374.

[17] J. Yang, R. Zhang, H. Zhao, et al., “Bioinspired Copper Single-Atom Nanozyme as a Superoxide Dismutase-Like Antioxidant for Sepsis Treatment,” Exploration 2 (2022): 20210267.

[18] a) L. Wu, H. Lin, X. Cao, et al., “Bioorthogonal Cu Single-Atom Nanozyme for Synergistic Nanocatalytic Therapy, Photothermal Therapy, Cuproptosis and Immunotherapy,” Angewandte Chemie, International Edition 63 (2024): e202405937. b) Y. Wang, V. K. Paidi, W. Wang, et al., “Spatial Engineering of Single-Atom Fe Adjacent to Cu-Assisted Nanozymes for Biomimetic O₂ Activation,” Nature Communications 15 (2024): 2239.

[19] a) Y. Li, R. Fu, Z. Duan, C. Zhu, and D. Fan, “Artificial Nonenzymatic Antioxidant MXene Nanosheet-Anchored Injectable Hydrogel as a Mild Photothermal-Controlled Oxygen Release Platform for Diabetic Wound Healing,” ACS Nano 16 (2022): 7486. b) H. Bao, Y. Qiu, X. Peng, et al., “Isolated Copper Single Sites for High-Performance Electroreduction of Carbon Monoxide to Multicarbon Products,” Nature Communications2021, 12, 238; c) Y. Zhu, Z. Wang, R. Zhao, et al., “Pt Decorated Ti₃C₂T_X MXene With NIR-II Light Amplified Nanozyme Catalytic Activity for Efficient Phototheranostics,” ACS Nano 16 (2022): 3105.

[20] a) L. Zhang, A. Song, Q.-C. Yang, et al., “Integration of AIEgens Into Covalent Organic Frameworks for Pyroptosis and Ferroptosis Primed Cancer Immunotherapy,” Nature Communications 14 (2023): 5355. b) J. Liu, J. Zhan, Y. Zhang, et al., “Ultrathin Clay Nanoparticles-Mediated Mutual Reinforcement of Ferroptosis and Cancer Immunotherapy,” Advanced Materials 36, no. 9 (2023): e2309562. c) T. Li, Y. Zhang, J. Zhu, et al., “A pH-Activatable Copper-Biomineralized Proenzyme for Synergistic Chemodynamic/Chemo-Immunotherapy Against Aggressive Cancers,” Advanced Materials 35 (2023): e2210201.

[21] Y. Tang, H. K. Bisoyi, X. M. Chen, et al., “Pyroptosis-Mediated Synergistic Photodynamic and Photothermal Immunotherapy Enabled by a Tumor-Membrane-Targeted Photosensitive Dimer,” Advanced Materials 35 (2023): e2300232.

[22] a) X. Zhang, Z. Wei, T. Yong, et al., “Cell Microparticles Loaded With Tumor Antigen and Resiquimod Reprogram Tumor-associated Macrophages and Promote Stem-Like CD8+ T Cells to Boost Anti-PD-1 Therapy,” Nature Communications 14 (2023): 5653. b) D. Jana, B. He, Y. Chen, J. Liu, and Y. Zhao, “A Defect-Engineered Nanozyme for Targeted NIR-II Photothermal Immunotherapy of Cancer,” Advanced Materials 36, no. 10 (2022): e2206401.

[23] H. Shan, J. Shi, T. Chen, et al., “Modulating Catalytic Activity and Stability of Atomically Precise Gold Nanoclusters as Peroxidase Mimics via Ligand Engineering,” ACS Nano 17 (2023): 2368.

[24] a) Y. Liu, J. Wu, Y. Jin, et al., “Copper(I) Phosphide Nanocrystals for In Situ Self-Generation Magnetic Resonance Imaging-Guided Photothermal-Enhanced Chemodynamic Synergetic Therapy Resisting Deep-Seated Tumor,” Advanced Functional Materials 29 (2019): 1904678. b) B. Ma, S. Wang, F. Liu, et al., “Self-Assembled Copper–Amino Acid Nanoparticles for In Situ Glutathione “and” H₂O₂ Sequentially Triggered Chemodynamic Therapy,” Journal of the American Chemical Society 141 (2019): 849.

[25] M. Zhang, D. Yang, C. Dong, et al., “Two-Dimensional MXene-Originated In Situ Nanosonosensitizer Generation for Augmented and Synergistic Sonodynamic Tumor Nanotherapy,” ACS Nano 16 (2022): 9938.

[26] Y. Du, R. Zhang, J. Yang, et al., “A “Closed-Loop” Therapeutic Strategy Based on Mutually Reinforced Ferroptosis and Immunotherapy,” Advanced Functional Materials 32, no. 13 (2022): 2111784.

[27] W. Chen, W. Xie, Z. Gao, et al., “Mild-Photothermal Effect Induced High Efficiency Ferroptosis-Boosted-Cuproptosis Based on Cu₂ O@Mn 3 Cu 3 O 8 Nanozyme,” Advancement of Science 10 (2023): e2303694.

[28] J. Ye, K. Zhang, X. Yang, et al., “Embedding Atomically Dispersed Manganese/Gadolinium Dual Sites in Oxygen Vacancy-Enriched Biodegradable Bimetallic Silicate Nanoplatform for Potentiating Catalytic Therapy,” Advancement of Science 11, no. 4 (2023): 2307424.

[29] C. Yang, M. Wang, M. Chang, M. Yuan, W. Zhang, J. Tan, B. Ding, P. Ma, and J. Lin, “Heterostructural Nanoadjuvant CuSe/CoSe₂ for Potentiating Ferroptosis and Photoimmunotherapy Through Intratumoral Blocked Lactate Efflux,” Journal of the American Chemical Society 145 (2023): 7205.

[30] L. Qiao, G. Zhu, T. Jiang, et al., “Self-Destructive Copper Carriers Induce Pyroptosis and Cuproptosis for Efficient Tumor Immunotherapy Against Dormant and Recurrent Tumors,” Advanced Materials 36, no. 8 (2023): e2308241.

[31] P. Zhu, Y. Pu, M. Wang, et al., “MnOOH-Catalyzed Autoxidation of Glutathione for Reactive Oxygen Species Production and Nanocatalytic Tumor Innate Immunotherapy,” Journal of the American Chemical Society 145 (2023): 5803.

[32] a) H. Yuan, P. Xia, X. Sun, et al., “Photothermal Nanozymatic Nanoparticles Induce Ferroptosis and Apoptosis Through Tumor Microenvironment Manipulation for Cancer Therapy,” Small 18 (2022): e2202161. b) J. Li, L. Zeng, Z. Wang, et al., “Cycloruthenated Self-Assembly With Metabolic Inhibition to Efficiently Overcome Multidrug Resistance in Cancers,” Advanced Materials 34 (2022): 2100245.

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2025 The Author(s). Exploration published by Henan University and John Wiley & Sons Australia, Ltd.