Oral proton-reprogrammed nanomedicine to break the inflammatory bowel disease-Clostridium difficile infection vicious cycle

Abstract

Clostridium difficile infection (CDI) is a major complication of inflammatory bowel disease (IBD), but the complicated treatment strategy of antimicrobial drugs and immunosuppressive drugs cannot effectively control the syndrome of IBD and CDI. Glycolysis in intestinal epithelial cells (IECs) leads to excessive production of lactic acid, which is considered to be a key factor in the destruction of the intestinal barrier by toxin B of Clostridium difficile (TcdB). In this study, we revealed that uncoupling protein 2 (UCP2) was highly expressed in IECs, and disrupted mitochondrial function to reprogram metabolism toward glycolysis, thereby weakening the resistance of IECs to TcdB. We developed an oral proton-reprogrammed nanomedicine (OPR) to convert the metabolism of IECs from glycolysis to oxidative phosphorylation, which can effectively alleviate IBD and CDI syndromes. OPR blocks the rapid delivery of protons to the mitochondrial matrix induced by UCP2, which restores mitochondrial membrane potential (MMP) and drives protons to reenter the matrix through ATP synthase, mediating the recovery of mitochondrial function. In mouse IBD and IBD complicated CDI (IBD-CDI) models, OPR selectively targeted inflammatory colonic lesions, alleviated mitochondria-dependent pyroptosis of IECs, inhibited macrophage STING pathway activation, and reduced lactic acid levels. Especially in the treatment of IBD-CDI syndrome, the therapeutic effect of OPR is far better than the combination of antibiotics and immunosuppressive drugs. In summary, our study establishes UCP2-mediated mitochondrial disability as a key mechanism for the functioning of TcdB and suggests that IEC metabolic reprogramming could be a therapeutic target.

Keywords

Clostridium difficile infection / inflammatory bowel disease / metabolic reprogramming / nanomedicine / uncoupling protein 2

References

[1] Solitano, Virginia, Charles N. Bernstein, Iris Dotan, Axel Dignass, Rhondell Domilici, Marla C. Dubinsky, Richard B. Gearry, et al. 2025. “Shaping the Future of Inflammatory Bowel Disease: A Global Research Agenda for Better Management and Public Health Response.” Nature Reviews Gastroenterology & Hepatology 22: 438–52. https://doi.org/10.1038/s41575-025-01063-x

[2] Hanada, Yuri, Sahil Khanna, Edward V. Loftus, Laura E. Raffals, and Darrell S. Pardi. 2018. “Non-Clostridium Difficile Bacterial Infections Are Rare in Patients With Flares of Inflammatory Bowel Disease.” Clinical Gastroenterology and Hepatology 16: 528–33. https://doi.org/10.1016/j.cgh.2017.10.008

[3] Singh, Harminder, Zoann Nugent, B. Nancy Yu, Lisa M. Lix, Laura E. Targownik, and Charles N. Bernstein. 2017. “Higher Incidence of Clostridium difficile Infection Among Individuals With Inflammatory Bowel Disease.” Gastroenterology 153: 430–8.e2. https://doi.org/10.1053/j.gastro.2017.04.044

[4] Vaughn, Byron P., Alexander Khoruts, and Monika Fischer. 2025. “Diagnosis and Management of Clostridioides difficile in Inflammatory Bowel Disease.” American Journal of Gastroenterology 120: 313–9. https://doi.org/10.14309/ajg.0000000000003076

[5] Kordus, Shannon L., Audrey K. Thomas, and D. Borden Lacy. 2022. “Clostridioides Difficile Toxins: Mechanisms of Action and Antitoxin Therapeutics.” Nature Reviews Microbiology 20: 285–98. https://doi.org/10.1038/s41579-021-00660-2

[6] Zhang, Ting, Qian-Yun Lin, Jia-Xi Fei, Yan Zhang, Min-Yi Lin, Shuang-Hong Jiang, Pu Wang, and Ye Chen. 2016. “Clostridium Difficile Infection Worsen Outcome of Hospitalized Patients with Inflammatory Bowel Disease.” Scientific Reports 6: 29791. https://doi.org/10.1038/srep29791

[7] Del Vecchio, Livio Enrico, Marcello Fiorani, Ege Tohumcu, Stefano Bibbò, Serena Porcari, Maria Cristina Mele, Marco Pizzoferrato, et al. 2022. “Risk Factors, Diagnosis, and Management of Clostridioides difficile Infection in Patients with Inflammatory Bowel Disease.” Microorganisms 10: 1315. https://doi.org/10.3390/microorganisms10071315

[8] Jin, Xi, Dong Chen, Ruo-Heng Zheng, Hong Zhang, Yi-Peng Chen, and Zun Xiang. 2017. “miRNA-133a-UCP2 Pathway Regulates Inflammatory Bowel Disease Progress by Influencing Inflammation, Oxidative Stress and Energy Metabolism.” World Journal of Gastroenterology 23: 76–86. https://doi.org/10.3748/wjg.v23.i1.76

[9] Brand, Martin D., and Telma C. Esteves. 2005. “Physiological Functions of the Mitochondrial Uncoupling Proteins UCP2 and UCP3.” Cell Metabolism 2: 85–93. https://doi.org/10.1016/j.cmet.2005.06.002

[10] Vozza, Angelo, Giovanni Parisi, Francesco De Leonardis, Francesco M. Lasorsa, Alessandra Castegna, Daniela Amorese, Raffaele Marmo, et al. 2014. “UCP2 Transports C4 Metabolites out of Mitochondria, Regulating Glucose and Glutamine Oxidation.” Proceedings of the National Academy of Sciences 111: 960–5. https://doi.org/10.1073/pnas.1317400111

[11] Doherty, Joanne R., and John L. Cleveland. 2013. “Targeting Lactate Metabolism for Cancer Therapeutics.” Journal of Clinical Investigation 123: 3685–92. https://doi.org/10.1172/JCI69741

[12] Halestrap, Andrew P. 2013. “The SLC16 Gene Family – Structure, Role and Regulation in Health and Disease.” Molecular Aspects of Medicine 34: 337–49. https://doi.org/10.1016/j.mam.2012.05.003

[13] Vernia, Piero, Renzo Caprilli, Giovanni Latella, Fabrizio Barbetti, Fabio M. Magliocca, and Mauro Cittadini. 1988. “Fecal Lactate and Ulcerative Colitis.” Gastroenterology 95: 1564–8. https://doi.org/10.1016/S0016-5085(88)80078-7

[14] Tam, John, Therwa Hamza, Bing Ma, Kevin Chen, Greg L. Beilhartz, Jacques Ravel, Hanping Feng, and Roman A. Melnyk. 2018. “Host-Targeted Niclosamide Inhibits C. difficile Virulence and Prevents Disease in Mice Without Disrupting the Gut Microbiota.” Nature Communications 9: 5233. https://doi.org/10.1038/s41467-018-07705-w

[15] Chen, Yi, Mingju Shui, Qin Yuan, Chi Teng Vong, Zhengming Yang, Zhejie Chen, and Shengpeng Wang. 2023. “Wielding the Double-Edged Sword: Redox Drug Delivery Systems for Inflammatory Bowel Disease.” Journal of Controlled Release 358: 510–40. https://doi.org/10.1016/j.jconrel.2023.05.007

[16] Pecqueur, Claire, Marie-Clotilde Alves-Guerra, Chantal Gelly, Corinne Lévi-Meyrueis, Elodie Couplan, Sheila Collins, Daniel Ricquier, Frédéric Bouillaud, and Bruno Miroux. 2001. “Uncoupling Protein 2, in Vivo Distribution, Induction Upon Oxidative Stress, and Evidence for Translational Regulation*.” Journal of Biological Chemistry 276: 8705–12. https://doi.org/10.1074/jbc.M006938200

[17] Zhu, Xinyi, Henghui Li, Liuzhu Zhou, Huijun Jiang, Minghui Ji, and Jin Chen. 2023. “Evaluation of the Gut Microbiome Alterations in Healthy Rats After Dietary Exposure to Different Synthetic ZnO Nanoparticles.” Life Sciences 312: 121250. https://doi.org/10.1016/j.lfs.2022.121250

[18] Zhang, Chen-Yu, Laura E. Parton, Chian Ping Ye, Stefan Krauss, Ruichao Shen, Cheng-Ting Lin, John A. Porco, and Bradford B. Lowell. 2006. “Genipin Inhibits UCP2-Mediated Proton Leak and Acutely Reverses Obesity- and High Glucose-Induced β Cell Dysfunction in Isolated Pancreatic Islets.” Cell Metabolism 3: 417–27. https://doi.org/10.1016/j.cmet.2006.04.010

[19] Neves, Maria Isabel Landim, Alberto Valdés, Eric Keven Silva, Maria Angela A. Meireles, Elena Ibáñez, and Alejandro Cifuentes. 2022. “Study of the Reaction Between Genipin and Amino Acids, Dairy Proteins, and Milk to Form a Blue Colorant Ingredient.” Food Research International 157: 111240. https://doi.org/10.1016/j.foodres.2022.111240

[20] Pizarro, Fernando, Manuel Olivares, Eva Hertrampf, Dora I. Mazariegos, Miguel Arredondo, Angélica Letelier, and Virginia Gidi. 2002. “Iron Bis-Glycine Chelate Competes for the Nonheme-Iron Absorption Pathway.” The American Journal of Clinical Nutrition 76: 577–81. https://doi.org/10.1093/ajcn/76.3.577

[21] Ahmed, Rizwan, Noor ul ain Hira, Mingwei Wang, Shahid Iqbal, Jiang Yi, and Yacine Hemar. 2024. “Genipin, a Natural Blue Colorant Precursor: Source, Extraction, Properties, and Applications.” Food Chemistry 434: 137498. https://doi.org/10.1016/j.foodchem.2023.137498

[22] Chassaing, Benoit, Jesse D. Aitken, Madhu Malleshappa, and Matam Vijay-Kumar. 2014. “Dextran Sulfate Sodium (DSS)-Induced Colitis in Mice.” Current Protocols in Immunology 104: 1–14. https://doi.org/10.1002/0471142735.im1525s104

[23] Luis, Ana S., and Gunnar C. Hansson. 2023. “Intestinal Mucus and Their Glycans: A Habitat for Thriving Microbiota.” Cell Host & Microbe 31: 1087–100. https://doi.org/10.1016/j.chom.2023.05.026

[24] Kudelka, Matthew R., Sean R. Stowell, Richard D. Cummings, and Andrew S. Neish. 2020. “Intestinal Epithelial Glycosylation in Homeostasis and Gut Microbiota Interactions in IBD.” Nature Reviews Gastroenterology & Hepatology 17: 597–617. https://doi.org/10.1038/s41575-020-0331-7

[25] Neurath, Markus F., David Artis, and Christoph Becker. 2025. “The Intestinal Barrier: A Pivotal Role in Health, Inflammation, and Cancer.” The Lancet Gastroenterology & Hepatology 10: 573–92. https://doi.org/10.1016/S2468-1253(24)00390-X

[26] Li, Ruoyao, Yuanyuan Fan, Lulu Liu, Hongna Ma, Deyan Gong, Zhaohua Miao, Hua Wang, and Zhengbao Zha. 2022. “Ultrathin Hafnium Disulfide Atomic Crystals with ROS-Scavenging and Colon-Targeting Capabilities for Inflammatory Bowel Disease Treatment.” ACS Nano 16: 15026–41. https://doi.org/10.1021/acsnano.2c06151

[27] Song, Xiangping, Qiong Huang, Yuqi Yang, Liang Ma, Wenguang Liu, Chunlin Ou, Qiaohui Chen, et al. 2023. “Efficient Therapy of Inflammatory Bowel Disease (IBD) with Highly Specific and Durable Targeted Ta2C Modified with Chondroitin Sulfate (TACS).” Advanced Materials 35: 2301585. https://doi.org/10.1002/adma.202301585

[28] Bu, Yingzi, Mohamed Dit Mady Traore, Luchen Zhang, Lu Wang, Zhongwei Liu, Hongxiang Hu, Meilin Wang, Chengyi Li, and Duxin Sun. 2023. “A Gastrointestinal Locally Activating Janus Kinase Inhibitor to Treat Ulcerative Colitis.” Journal of Biological Chemistry 299: 105467. https://doi.org/10.1016/j.jbc.2023.105467

[29] Lee, Yonghyun, Kohei Sugihara, Merritt G. Gillilland, Sangyong Jon, Nobuhiko Kamada, and James J. Moon. 2020. “Hyaluronic Acid–Bilirubin Nanomedicine for Targeted Modulation of Dysregulated Intestinal Barrier, Microbiome and Immune Responses in Colitis.” Nature Materials 19: 118–26. https://doi.org/10.1038/s41563-019-0462-9

[30] Xiao, Fangfei, Fang Dong, Xiaolu Li, Youran Li, Guangjun Yu, Zhanju Liu, Yizhong Wang, and Ting Zhang. 2022. “Bifidobacterium longum CECT 7894 Improves the Efficacy of Infliximab for DSS-Induced Colitis Via Regulating the Gut Microbiota and Bile Acid Metabolism.” Frontiers in Pharmacology 13: 2022. https://doi.org/10.3389/fphar.2022.902337

[31] Burridge, Keith, and Krister Wennerberg. 2004. “Rho and Rac Take Center Stage.” Cell 116: 167–79. https://doi.org/10.1016/S0092-8674(04)00003-0

[32] Vadlamudi, Ratna K., Feng Li, Liana Adam, Diep Nguyen, Yasutaka Ohta, Thomas P. Stossel, and Rakesh Kumar. 2002. “Filamin Is Essential in Actin Cytoskeletal Assembly Mediated by p21-activated Kinase 1.” Nature Cell Biology 4: 681–90. https://doi.org/10.1038/ncb838

[33] Khanna, Sahil, Andrea Shin, and Ciarán P. Kelly. 2017. “Management of Clostridium difficile Infection in Inflammatory Bowel Disease: Expert Review from the Clinical Practice Updates Committee of the AGA Institute.” Clinical Gastroenterology and Hepatology 15: 166–74. https://doi.org/10.1016/j.cgh.2016.10.024

[34] Ajami, N. J., J. L. Cope, M. C. Wong, J. F. Petrosino, and L. Chesnel. 2018. “Impact of Oral Fidaxomicin Administration on the Intestinal Microbiota and Susceptibility to Clostridium difficile Colonization in Mice.” Antimicrobial Agents and Chemotherapy 62. https://doi.org/10.1128/aac.02112-17

[35] Haque, Parsa S., Kapur Neeraj, Terrence A. Barrett, and Arianne L. Theiss. 2024. “Mitochondrial Function and Gastrointestinal Diseases.” Nature Reviews Gastroenterology & Hepatology 21: 537–55. https://doi.org/10.1038/s41575-024-00931-2

[36] Bhattacharyya, Asima, Ranajoy Chattopadhyay, Sankar Mitra, and Sheila E. Crowe. 2014. “Oxidative Stress: An Essential Factor in the Pathogenesis of Gastrointestinal Mucosal Diseases.” Physiological Reviews 94: 329–54. https://doi.org/10.1152/physrev.00040.2012

[37] Dimitrov, Georges, Bernhard Ryffel, Dieudonnée Togbe, and Valérie Quesniaux. 2025. “cGAS-STING DNA-Sensing in Inflammatory Bowel Diseases.” Trends in Molecular Medicine 31: 165–80. https://doi.org/10.1016/j.molmed.2024.10.002

[38] Privitera, Giuseppe, Nitish Rana, Alessandro Armuzzi, and Theresa T. Pizarro. 2023. “The Gasdermin Protein Family: Emerging Roles in Gastrointestinal Health and Disease.” Nature Reviews Gastroenterology & Hepatology 20: 366–87. https://doi.org/10.1038/s41575-023-00743-w

[39] Razik, Roshan, Amir Rumman, Zoya Bahreini, Allison McGeer, and Geoffrey C. Nguyen. 2016. “Recurrence of Clostridium difficile Infection in Patients with Inflammatory Bowel Disease: The RECIDIVISM Study.” American Journal of Gastroenterology 111: 1141–6. https://doi.org/10.1038/ajg.2016.187

[40] Song, Jung Hoon, and You Sun Kim. 2019. “Recurrent Clostridium difficile Infection: Risk Factors, Treatment, and Prevention.” Gut and Liver 13: 16–24. https://doi.org/10.5009/gnl18071

[41] Simeon, Rudo, Mengqiu Jiang, Ana M. Chamoun-Emanuelli, Hua Yu, Yongrong Zhang, Ran Meng, Zeyu Peng, et al. 2019. “Selection and Characterization of Ultrahigh Potency Designed Ankyrin Repeat Protein Inhibitors of C. difficile Toxin B.” PLoS Biology 17: e3000311. https://doi.org/10.1371/journal.pbio.3000311

[42] Aguilar, Esther, Pauline Esteves, Tiphaine Sancerni, Véronique Lenoir, Thomas Aparicio, Frédéric Bouillaud, Renaud Dentin, et al. 2019. “UCP2 Deficiency Increases Colon Tumorigenesis by Promoting Lipid Synthesis and Depleting NADPH for Antioxidant Defenses.” Cell Reports 28: 2306–16.e5. https://doi.org/10.1016/j.celrep.2019.07.097

[43] Lee, May Y., Andrea Leonardi, Thomas J. Begley, and J. Andrés Melendez. 2020. “Loss of Epitranscriptomic Control of Selenocysteine Utilization Engages Senescence and Mitochondrial Reprogramming.” Redox Biology 28: 101375. https://doi.org/10.1016/j.redox.2019.101375

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