Bacterial Biohybrids With Dual Magnet and Hypoxia Tropism for Ferroptosis Activation in Deep Tumor Regions

Abstract

Effectively delivering therapeutics to deep tumor regions is crucial for successful treatment but remains a formidable challenge. The severe hypoxia common in these areas can inhibit various therapeutic-induced cell death pathways, including ferroptosis. Herein, we present the design of a bacterial biohybrid (Ec@ZFOY) with dual magnetic and hypoxic tropism for targeted therapeutic delivery and ferroptosis activation in the deep tumor region. This biohybrid is constructed using hypoxia-targeted Escherichia coli and magnetic Zn_0.16Fe_1.24O₄ (ZFO) nanoparticles loaded with YC-1, a hypoxia-inducible factor-1α (HIF-1α) inhibitor. ZFO exhibits peroxidase- and glutathione oxidase-mimetic activities, catalyzing tumor-derived H₂O₂ into hydroxyl radical and inhibiting glutathione peroxidase 4, thereby inducing ferroptosis. Additionally, Ec@ZFOY demonstrates pH-responsive YC-1 release, which inhibits HIF-1α and reduces lipid droplets, enhancing ferroptosis through the release of polyunsaturated fatty acids. Both in vitro and in vivo experiments confirm the significant therapeutic efficacy of Ec@ZFOY. This study unveils the design of magnetically and hypoxia-tropic bacterial biohybrids for activating ferroptosis in deep tumor sites, highlighting their potential for other therapeutic delivery and disease modulation applications.

Keywords

bacterial biohybrids / ferroptosis / hypoxia / tumor penetration

References

[1] J. Hahn, S. Ding, J. Im, T. Harimoto, K. W. Leong, and T. Danino, “Bacterial Therapies at the Interface of Synthetic Biology and Nanomedicine,” Nature Reviews Bioengineering 2 (2024): 120–135, https://doi.org/10.1038/s44222-023-00119-4.

[2] C. R. Gurbatri, N. Arpaia, and T. Danino, “Engineering Bacteria as Interactive Cancer Therapies,” Science 378 (2022): 858–864, https://doi.org/10.1126/science.add9667.

[3] A. Cubillos-Ruiz, T. Guo, A. Sokolovska, et al., “Engineering Living Therapeutics With Synthetic Biology,” Nature Reviews Drug Discovery 20 (2021): 941–960, https://doi.org/10.1038/s41573-021-00285-3.

[4] T. He, Z. Zhao, Z. Luo, et al., “Advances in Microbial Decorations and Its Applications in Drug Delivery,” Acta Materia Medica 2 (2023): 466–479.

[5] S. Zhu, Y. Cheng, J. Wang, et al., “Biohybrid Magnetic Microrobots: An Intriguing and Promising Platform in Biomedicine,” Acta Biomaterialia 169 (2023): 88–106.

[6] Z. Hosseinidoust, B. Mostaghaci, O. Yasa, B. W. Park, A. V. Singh, and M. Sitti, “Bioengineered and Biohybrid Bacteria-Based Systems for Drug Delivery,” Advanced Drug Delivery Reviews 106 (2016): 27–44, https://doi.org/10.1016/j.addr.2016.09.007.

[7] D. Akin, J. Sturgis, K. Ragheb, et al., “Bacteria-Mediated Delivery of Nanoparticles and Cargo Into Cells,” Nature Nanotechnology 2 (2007): 441, https://doi.org/10.1038/nnano.2007.149.

[8] C.-H. Luo, C.-T. Huang, C.-H. Su, and C.-S. Yeh, “Bacteria-Mediated Hypoxia-Specific Delivery of Nanoparticles for Tumors Imaging and Therapy,” Nano Letters 16 (2016): 3493–3499, https://doi.org/10.1021/acs.nanolett.6b00262.

[9] M. Huo, L. Wang, Y. Chen, and J. Shi, “Nanomaterials/Microorganism-Integrated Microbiotic Nanomedicine,” Nano Today 32 (2020): 100854, https://doi.org/10.1016/j.nantod.2020.100854.

[10] Y. X. Zhu, H. R. Jia, Y. W. Jiang, et al., “A Red Blood Cell-Derived Bionic Microrobot Capable of Hierarchically Adapting to Five Critical Stages in Systemic Drug Delivery,” Exploration (2024): 20230105, https://doi.org/10.1002/EXP.20230105.

[11] X. Zeng, P. Li, S. Yan, and B.-F. Liu, “Reduction/pH-Responsive Disassemblable MOF-Microbial Nanohybrid for Targeted Tumor Penetration and Synergistic Therapy,” Chemical Engineering Journal 452 (2023): 139517, https://doi.org/10.1016/j.cej.2022.139517.

[12] X. Huang, H. Guo, L. Wang, and Z. Shao, “Engineered Microorganism-Based Delivery Systems for Targeted Cancer Therapy: A Narrative Review,” Biomaterials Translational 3 (2022): 201.

[13] Y. Li, Q. Leng, Y. Zhang, et al., “Anaerobic Bacteria Mediated ‘Smart Missile’ Targeting Tumor Hypoxic Area Enhances the Therapeutic Outcome of Lung Cancer,” Chemical Engineering Journal 438 (2022): 135566, https://doi.org/10.1016/j.cej.2022.135566.

[14] Z. Y. Han, C. Zhang, J. X. An, et al., “Metabolic Regulation of Tumor Microenvironment With Biohybrid Bacterial Bioreactor for Enhanced Cancer Chemo-Immunotherapy,” Advanced Functional Materials 33, no. 35 (2023): 2302728, https://doi.org/10.1002/adfm.202302728.

[15] Y. Ruan, H. Zhuang, X. Zeng, et al., “Engineered Microbial Nanohybrids for Tumor-Mediated NIR II Photothermal Enhanced Ferroptosis/Cuproptosis and Immunotherapy,” Advanced Healthcare Materials 13 (2024): 2302537, https://doi.org/10.1002/adhm.202302537.

[16] S. Yan, X. Zeng, Y. Wang, and B. F. Liu, “Biomineralization of Bacteria by a Metal–Organic Framework for Therapeutic Delivery,” Advanced Healthcare Materials 9 (2020): 2000046, https://doi.org/10.1002/adhm.202000046.

[17] V. M. Moreno, E. Álvarez, I. Izquierdo-Barba, A. Baeza, J. Serrano-López, and M. Vallet-Regí, “Bacteria as Nanoparticles Carrier for Enhancing Penetration in a Tumoral Matrix Model,” Advanced Materials Interfaces 7 (2020): 1901942, https://doi.org/10.1002/admi.201901942.

[18] B. He, X. Sui, B. Yu, S. Wang, Y. Shen, and H. Cong, “Recent Advances in Drug Delivery Systems for Enhancing Drug Penetration Into Tumors,” Drug Delivery 27 (2020): 1474–1490, https://doi.org/10.1080/10717544.2020.1831106.

[19] J. Ding, J. Chen, L. Gao, et al., “Engineered Nanomedicines With Enhanced Tumor Penetration,” Nano Today 29 (2019): 100800, https://doi.org/10.1016/j.nantod.2019.100800.

[20] X. Zhang, X. Chen, J. Song, J. Zhang, X. Ren, and Y. Zhao, “Size-Transformable Nanostructures: From Design to Biomedical Applications,” Advanced Materials 32 (2020): 2003752, https://doi.org/10.1002/adma.202003752.

[21] G. Huang, Y. Qiu, F. Yang, et al., “Magnetothermally Triggered Free-Radical Generation for Deep-Seated Tumor Treatment,” Nano Letters 21 (2021): 2926–2931, https://doi.org/10.1021/acs.nanolett.1c00009.

[22] J. Zhou, T. Schmid, S. Schnitzer, and B. Brüne, “Tumor Hypoxia and Cancer Progression,” Cancer Letters 237 (2006): 10–21, https://doi.org/10.1016/j.canlet.2005.05.028.

[23] X. Lou, Z. Chen, Z. He, M. Sun, and J. Sun, “Bacteria-Mediated Synergistic Cancer Therapy: Small Microbiome Has a Big Hope,” Nano-Micro Letters 13 (2021): 1–26, https://doi.org/10.1007/s40820-020-00560-9.

[24] C. K. Schmidt, M. Medina-Sánchez, R. J. Edmondson, and O. G. Schmidt, “Engineering Microrobots for Targeted Cancer Therapies From a Medical Perspective,” Nature Communications 11 (2020): 1–18.

[25] L. Guo, H. Chen, J. Ding, P. Rong, M. Sun, and W. Zhou, “Surface Engineering Salmonella With pH-Responsive Polyserotonin and Self-Activated DNAzyme for Better Microbial Therapy of Tumor,” Exploration (2023): 20230017, https://doi.org/10.1002/EXP.20230017.

[26] M. B. Akolpoglu, Y. Alapan, N. O. Dogan, et al., “Magnetically Steerable Bacterial Microrobots Moving in 3D Biological Matrices for Stimuli-Responsive Cargo Delivery,” Science Advances 8 (2022): eabo6163, https://doi.org/10.1126/sciadv.abo6163.

[27] J. F. Toso, V. J. Gill, P. Hwu, et al., “Phase I Study of the Intravenous Administration of Attenuated Salmonella Typhimurium to Patients with Metastatic Melanoma,” Journal of Clinical Oncology 20 (2002): 142–152, https://doi.org/10.1200/JCO.2002.20.1.142.

[28] P. Basu, A. Mehta, M. Jain, et al., “A Randomized Phase 2 Study of ADXS11-001 Listeria Monocy'togenes-Listeriolysin O Immunotherapy With or Without Cisplatin in Treatment of Advanced Cervical Cancer,” International Journal of Gynecological Cancer 28 (2018): 764–772, https://doi.org/10.1097/IGC.0000000000001235.

[29] J. M. Brown, “Tumor Hypoxia in Cancer Therapy,” Methods in Enzymology 435 (2007): 295–321.

[30] T. Hompland, C. S. Fjeldbo, and H. Lyng, “Tumor Hypoxia as a Barrier in Cancer Therapy: Why Levels Matter,” Cancers (Basel) 13 (2021): 499, https://doi.org/10.3390/cancers13030499.

[31] P. Carmeliet, Y. Dor, J.-M. Herbert, et al., “Role of HIF-1α in Hypoxia-Mediated Apoptosis, Cell Proliferation and Tumor Angiogenesis,” Nature 394 (1998): 485–490, https://doi.org/10.1038/28867.

[32] P. Tsvetkov, S. Coy, B. Petrova, et al., “Copper Induces Cell Death by Targeting Lipoylated TCA Cycle Proteins,” Science 375 (2022): 1254–1261, https://doi.org/10.1126/science.abf0529.

[33] D. C. Fuhrmann, A. Mondorf, J. Beifuß, M. Jung, and B. Brüne, “Hypoxia Inhibits Ferritinophagy, Increases Mitochondrial Ferritin, and Protects From Ferroptosis,” Redox Biology 36 (2020): 101670, https://doi.org/10.1016/j.redox.2020.101670.

[34] W. S. Yang and B. R. Stockwell, “Ferroptosis: Death by Lipid Peroxidation,” Trends in Cell Biology 26 (2016): 165–176, https://doi.org/10.1016/j.tcb.2015.10.014.

[35] X. Jiang, B. R. Stockwell, and M. Conrad, “Ferroptosis: Mechanisms, Biology and Role in Disease,” Nature Reviews Molecular Cell Biology 22 (2021): 266–282, https://doi.org/10.1038/s41580-020-00324-8.

[36] G. Lei, L. Zhuang, and B. Gan, “Targeting Ferroptosis as a Vulnerability in Cancer,” Nature Reviews Cancer 22 (2022): 381–396, https://doi.org/10.1038/s41568-022-00459-0.

[37] X. Chen, R. Kang, G. Kroemer, and D. Tang, “Broadening Horizons: The Role of Ferroptosis in Cancer,” Nature Reviews Clinical Oncology 18 (2021): 280–296, https://doi.org/10.1038/s41571-020-00462-0.

[38] S. Guo, W. Xiong, J. Zhu, et al. “A STING Pathway-Activatable Contrast Agent for MRI-Guided Tumor Immunoferroptosis Synergistic Therapy,” Biomaterials 302 (2023): 122300.

[39] Q. Fan, W. Xiong, H. Zhou, et al., “Achieving Practical High-Energy-Density Lithium-Metal Batteries by a Dual-Anion Regulated Electrolyte,” Advanced Materials 35 (2023): 2301171.

[40] S. J. Dixon and B. R. Stockwell, “The Hallmarks of Ferroptosis,” Annual Review of Cancer Biology 3 (2019): 35–54, https://doi.org/10.1146/annurev-cancerbio-030518-055844.

[41] I. Mylonis, G. Simos, and E. Paraskeva, “Hypoxia-Inducible Factors and the Regulation of Lipid Metabolism,” Cells 8 (2019): 214, https://doi.org/10.3390/cells8030214.

[42] K. Bensaad, E. Favaro, C. A. Lewis, et al., “Fatty Acid Uptake and Lipid Storage Induced by HIF-1α Contribute to Cell Growth and Survival After Hypoxia-Reoxygenation,” Cell Reports 9 (2014): 349–365, https://doi.org/10.1016/j.celrep.2014.08.056.

[43] L. A. Broadfield, A. A. Pane, A. Talebi, J. V. Swinnen, and S.-M. Fendt, “Lipid Metabolism in Cancer: New Perspectives and Emerging Mechanisms,” Developmental Cell 56 (2021): 1363–1393, https://doi.org/10.1016/j.devcel.2021.04.013.

[44] Z. Gao, S. Zheng, K.-I. Kamei, and C. Tian, “Recent Progress in Cancer Therapy Based on the Combination of Ferroptosis with Photodynamic Therapy,” Acta Materia Medica 1 (2022): 411–426.

[45] G. Tang, J. He, J. Liu, X. Yan, and K. Fan, “Nanozyme for Tumor Therapy: Surface Modification Matters,” Exploration (2021): 75–89, https://doi.org/10.1002/EXP.20210005.

[46] Z. Dong, P. Liang, G. Guan, et al., “Overcoming Hypoxia-Induced Ferroptosis Resistance via a 19 F/ 1 H-MRI Traceable Core-Shell Nanostructure,” Angewandte Chemie International Edition 61 (2022): e202206074, https://doi.org/10.1002/anie.202206074.

[47] H. Yang, X. Yao, Y. Liu, X. Shen, M. Li, and Z. Luo, “Ferroptosis Nanomedicine: Clinical Challenges and Opportunities for Modulating Tumor Metabolic and Immunological Landscape,” ACS Nano 17 (2023): 15328–15353, https://doi.org/10.1021/acsnano.3c04632.

[48] M. Hou, M. Liu, H. Yu, et al., “Spatially Asymmetric Nanoparticles for Boosting Ferroptosis in Tumor Therapy,” Nano Letters 24 (2024): 1284–1293, https://doi.org/10.1021/acs.nanolett.3c04293.

[49] Y. Zhu, P. Gong, J. Wang, et al., “Amplification of Lipid Peroxidation by Regulating Cell Membrane Unsaturation To Enhance Chemodynamic Therapy,” Angewandte Chemie 135 (2023): e202218407, https://doi.org/10.1002/ange.202218407.

[50] L. Huang, J. Zhu, W. Xiong, et al., “Tumor-Generated Reactive Oxygen Species Storm for High-Performance Ferroptosis Therapy,” ACS Nano 17 (2023): 11492–11506, https://doi.org/10.1021/acsnano.3c01369.

[51] Y. Wang, S. Li, X. Wang, et al., “Smart Transformable Nanomedicines for Cancer Therapy,” Biomaterials (2021): 120737, https://doi.org/10.1016/j.biomaterials.2021.120737.

[52] N. Mirkhani, M. G. Christiansen, T. Gwisai, S. Menghini, and S. Schuerle, “Spatially Selective Delivery of Living Magnetic Microrobots Through Torque-focusing,” Nature Communications 15 (2024): 2160, https://doi.org/10.1038/s41467-024-46407-4.

[53] T. Gwisai, N. Mirkhani, M. G. Christiansen, T. T. Nguyen, V. Ling, and S. Schuerle, “Magnetic Torque-Driven Living Microrobots for Increased Tumor Infiltration,” Science Robotics 7 (2022): eabo0665, https://doi.org/10.1126/scirobotics.abo0665.

[54] P. Xue, H. Zhuang, T. Bai, et al., “Iron (II)-Based Metal-Organic Framework Nanozyme for Boosting Tumor Ferroptosis Through Inhibiting DNA Damage Repair and System Xc,” Journal of Nanbiotechnology 22 (2024): 228, https://doi.org/10.1186/s12951-024-02508-2.

[55] A. G. Kolhatkar, A. C. Jamison, D. Litvinov, R. C. Willson, and T. R. Lee, “Tuning the Magnetic Properties of Nanoparticles,” International Journal of Molecular Sciences 14 (2013): 15977–16009, https://doi.org/10.3390/ijms140815977.

[56] L. Huang, J. Zhu, G. Wu, et al., “A Strategy of “Adding Fuel to the Flames” Enables a Self-accelerating Cycle of Ferroptosis-Cuproptosis for Potent Antitumor Therapy,” Biomaterials 311 (2024): 122701.

[57] W. Wang, Y. Cui, X. Wei, et al., “CuCo₂O₄ Nanoflowers With Multiple Enzyme Activities for Treating Bacterium-Infected Wounds via Cuproptosis-Like Death,” ACS Nano 18 (2024): 15845–15863, https://doi.org/10.1021/acsnano.4c02825.

[58] J. Shan, X. Jin, C. Zhang, et al., “Metal Natural Product Complex Ru-Procyanidins With Quadruple Enzymatic Activity Combat Infections From Drug-Resistant Bacteria,” Acta Pharmaceutica Sinica B 14 (2024): 2298–2316, https://doi.org/10.1016/j.apsb.2023.12.017.

RIGHTS & PERMISSIONS

2026 The Author(s). Exploration published by Henan University and John Wiley & Sons Australia, Ltd.