Human Plasminogen Exacerbates Clostridioides difficile Enteric Disease and Alters the Spore Surface

Milena M. Awad, Melanie L. Hutton, Adam J. Quek, William P. Klare, Steven J. Mileto, Kate Mackin, Diane Ly, Viola Oorschot, Marijana Bosnjak, Grant Jenkin, Paul J. Conroy, Nick West, Alex Fulcher, Adam Costin, Christopher J. Day, Michael P. Jennings, Robert L. Medcalf, Martina Sanderson-Smith, Stuart J. Cordwell, Ruby H. P. LawJames C. Whisstock, Dena Lyras

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background & Aims The protease plasmin is an important wound healing factor, but it is not clear how it affects gastrointestinal infection–mediated damage, such as that resulting from Clostridioides difficile. We investigated the role of plasmin in C difficile–associated disease. This bacterium produces a spore form that is required for infection, so we also investigated the effects of plasmin on spores. Methods C57BL/6J mice expressing the precursor to plasmin, the zymogen human plasminogen (hPLG), or infused with hPLG were infected with C difficile, and disease progression was monitored. Gut tissues were collected, and cytokine production and tissue damage were analyzed by using proteomic and cytokine arrays. Antibodies that inhibit either hPLG activation or plasmin activity were developed and structurally characterized, and their effects were tested in mice. Spores were isolated from infected patients or mice and visualized using super-resolution microscopy; the functional consequences of hPLG binding to spores were determined. Results hPLG localized to the toxin-damaged gut, resulting in immune dysregulation with an increased abundance of cytokines (such as interleukin [IL] 1A, IL1B, IL3, IL10, IL12B, MCP1, MP1A, MP1B, GCSF, GMCSF, KC, TIMP-1), tissue degradation, and reduced survival. Administration of antibodies that inhibit plasminogen activation reduced disease severity in mice. C difficile spores bound specifically to hPLG and active plasmin and degraded their surface, facilitating rapid germination. Conclusions We found that hPLG is recruited to the damaged gut, exacerbating C difficile disease in mice. hPLG binds to C difficile spores, and, upon activation to plasmin, remodels the spore surface, facilitating rapid spore germination. Inhibitors of plasminogen activation might be developed for treatment of C difficile or other infection-mediated gastrointestinal diseases.
Original languageEnglish
JournalGastroenterology
DOIs
Publication statusAccepted/In press - 20 Jun 2020

Keywords

  • pathogenic bacteria
  • microbe
  • fibrinolytic system
  • liver enzyme

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Cite this

Awad, M. M., Hutton, M. L., Quek, A. J., Klare, W. P., Mileto, S. J., Mackin, K., Ly, D., Oorschot, V., Bosnjak, M., Jenkin, G., Conroy, P. J., West, N., Fulcher, A., Costin, A., Day, C. J., Jennings, M. P., Medcalf, R. L., Sanderson-Smith, M., Cordwell, S. J., ... Lyras, D. (Accepted/In press). Human Plasminogen Exacerbates Clostridioides difficile Enteric Disease and Alters the Spore Surface. Gastroenterology. https://doi.org/10.1053/j.gastro.2020.06.032