Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease

Steven J. Mileto; Thierry Jardé; Kevin O. Childress; Jaime L. Jensen; Ashleigh P. Rogers; Genevieve Kerr; Melanie L. Hutton; Michael J. Sheedlo; Sarah C. Bloch; John A. Shupe; Katja Horvay; Tracey Flores; Rebekah Engel; Simon Wilkins; Paul J. McMurrick; D. Borden Lacy; Helen E. Abud; Dena Lyras

doi:10.1073/pnas.1915255117

Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease

Steven J. Mileto, Thierry Jardé, Kevin O. Childress, Jaime L. Jensen, Ashleigh P. Rogers, Genevieve Kerr, Melanie L. Hutton, Michael J. Sheedlo, Sarah C. Bloch, John A. Shupe, Katja Horvay, Tracey Flores, Rebekah Engel, Simon Wilkins, Paul J. McMurrick, D. Borden Lacy, Helen E. Abud, Dena Lyras

Research output: Contribution to journal › Article › peer-review

60 Scopus citations

Abstract

Gastrointestinal infections often induce epithelial damage that must be repaired for optimal gut function. While intestinal stem cells are critical for this regeneration process [R. C. van der Wath, B. S. Gardiner, A. W. Burgess, D. W. Smith, PLoS One 8, e73204 (2013); S. Kozar et al., Cell Stem Cell 13, 626-633 (2013)], how they are impacted by enteric infections remains poorly defined. Here, we investigate infection-mediated damage to the colonic stem cell compartment and how this affects epithelial repair and recovery from infection. Using the pathogen Clostridioides difficile, we show that infection disrupts murine intestinal cellular organization and integrity deep into the epithelium, to expose the otherwise protected stem cell compartment, in a TcdB-mediated process. Exposure and susceptibility of colonic stem cells to intoxication compromises their function during infection, which diminishes their ability to repair the injured epithelium, shown by altered stem cell signaling and a reduction in the growth of colonic organoids from stem cells isolated from infected mice. We also show, using both mouse and human colonic organoids, that TcdB from epidemic ribotype 027 strains does not require Frizzled 1/2/7 binding to elicit this dysfunctional stem cell state. This stem cell dysfunction induces a significant delay in recovery and repair of the intestinal epithelium of up to 2 wk post the infection peak. Our results uncover a mechanism by which an enteric pathogen subverts repair processes by targeting stem cells during infection and preventing epithelial regeneration, which prolongs epithelial barrier impairment and creates an environment in which disease recurrence is likely.

Original language	English (US)
Pages (from-to)	8064-8073
Number of pages	10
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	117
Issue number	14
DOIs	https://doi.org/10.1073/pnas.1915255117
State	Published - Apr 7 2020
Externally published	Yes

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS. We thank the Monash Histology Platform, Department of Anatomy and Developmental Biology (Monash University, Clayton, Australia), Monash Biomedicine Discovery Institute Organoid Program, and Monash Micro Imaging (Monash University, Clayton, Australia) for their technical assistance with pathology and confocal imaging. We thank Jamie Rossjohn and Ben Kile for manuscript discussions, Kay Washington for pathology scoring, and Bliss Cunningham for assistance in animal handling. This work was supported by National Health and Medical Research Council Grant GNT1107812 (to D.L. and H.E.A.). Work in the D.B.L. laboratory was supported by NIH National Institute of Allergy and Infectious Diseases Grant AI95755, US Department of Veterans Affairs Grant BX002943, Molecular Biophysics Training Grant T32GM008320 (to K.O.C.), and Gastroenterology Training Grant T32DK007673 (to J.L.J. and M.J.S.). Collection of human tissue was supported by Monash Strategic Grant SGS15-0156 (to H.E.A., D.L., and P.J.M.) and, in part, by “Let’s Beat Bowel Cancer” (https://www.letsbeatbowelcancer.com.au/), a benevolent fundraising and public awareness foundation. This funder had no part in the design, conduct, outcomes, decision to publish, or the drafting of this manuscript.

Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.

Keywords

Clostridium difficile
Gut infection
Infection
Stem cell damage
Toxins

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Mileto, S. J., Jardé, T., Childress, K. O., Jensen, J. L., Rogers, A. P., Kerr, G., Hutton, M. L., Sheedlo, M. J., Bloch, S. C., Shupe, J. A., Horvay, K., Flores, T., Engel, R., Wilkins, S., McMurrick, P. J., Lacy, D. B., Abud, H. E., & Lyras, D. (2020). Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease. Proceedings of the National Academy of Sciences of the United States of America, 117(14), 8064-8073. https://doi.org/10.1073/pnas.1915255117

Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease. / Mileto, Steven J.; Jardé, Thierry; Childress, Kevin O. et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, No. 14, 07.04.2020, p. 8064-8073.

Research output: Contribution to journal › Article › peer-review

Mileto, SJ, Jardé, T, Childress, KO, Jensen, JL, Rogers, AP, Kerr, G, Hutton, ML, Sheedlo, MJ, Bloch, SC, Shupe, JA, Horvay, K, Flores, T, Engel, R, Wilkins, S, McMurrick, PJ, Lacy, DB, Abud, HE & Lyras, D 2020, 'Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 14, pp. 8064-8073. https://doi.org/10.1073/pnas.1915255117

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abstract = "Gastrointestinal infections often induce epithelial damage that must be repaired for optimal gut function. While intestinal stem cells are critical for this regeneration process [R. C. van der Wath, B. S. Gardiner, A. W. Burgess, D. W. Smith, PLoS One 8, e73204 (2013); S. Kozar et al., Cell Stem Cell 13, 626-633 (2013)], how they are impacted by enteric infections remains poorly defined. Here, we investigate infection-mediated damage to the colonic stem cell compartment and how this affects epithelial repair and recovery from infection. Using the pathogen Clostridioides difficile, we show that infection disrupts murine intestinal cellular organization and integrity deep into the epithelium, to expose the otherwise protected stem cell compartment, in a TcdB-mediated process. Exposure and susceptibility of colonic stem cells to intoxication compromises their function during infection, which diminishes their ability to repair the injured epithelium, shown by altered stem cell signaling and a reduction in the growth of colonic organoids from stem cells isolated from infected mice. We also show, using both mouse and human colonic organoids, that TcdB from epidemic ribotype 027 strains does not require Frizzled 1/2/7 binding to elicit this dysfunctional stem cell state. This stem cell dysfunction induces a significant delay in recovery and repair of the intestinal epithelium of up to 2 wk post the infection peak. Our results uncover a mechanism by which an enteric pathogen subverts repair processes by targeting stem cells during infection and preventing epithelial regeneration, which prolongs epithelial barrier impairment and creates an environment in which disease recurrence is likely.",

keywords = "Clostridium difficile, Gut infection, Infection, Stem cell damage, Toxins",

author = "Mileto, {Steven J.} and Thierry Jard{\'e} and Childress, {Kevin O.} and Jensen, {Jaime L.} and Rogers, {Ashleigh P.} and Genevieve Kerr and Hutton, {Melanie L.} and Sheedlo, {Michael J.} and Bloch, {Sarah C.} and Shupe, {John A.} and Katja Horvay and Tracey Flores and Rebekah Engel and Simon Wilkins and McMurrick, {Paul J.} and Lacy, {D. Borden} and Abud, {Helen E.} and Dena Lyras",

note = "Funding Information: ACKNOWLEDGMENTS. We thank the Monash Histology Platform, Department of Anatomy and Developmental Biology (Monash University, Clayton, Australia), Monash Biomedicine Discovery Institute Organoid Program, and Monash Micro Imaging (Monash University, Clayton, Australia) for their technical assistance with pathology and confocal imaging. We thank Jamie Rossjohn and Ben Kile for manuscript discussions, Kay Washington for pathology scoring, and Bliss Cunningham for assistance in animal handling. This work was supported by National Health and Medical Research Council Grant GNT1107812 (to D.L. and H.E.A.). Work in the D.B.L. laboratory was supported by NIH National Institute of Allergy and Infectious Diseases Grant AI95755, US Department of Veterans Affairs Grant BX002943, Molecular Biophysics Training Grant T32GM008320 (to K.O.C.), and Gastroenterology Training Grant T32DK007673 (to J.L.J. and M.J.S.). Collection of human tissue was supported by Monash Strategic Grant SGS15-0156 (to H.E.A., D.L., and P.J.M.) and, in part, by “Let{\textquoteright}s Beat Bowel Cancer” (https://www.letsbeatbowelcancer.com.au/), a benevolent fundraising and public awareness foundation. This funder had no part in the design, conduct, outcomes, decision to publish, or the drafting of this manuscript. Publisher Copyright: {\textcopyright} 2020 National Academy of Sciences. All rights reserved.",

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AU - Jensen, Jaime L.

AU - Rogers, Ashleigh P.

AU - Kerr, Genevieve

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AU - Shupe, John A.

AU - Horvay, Katja

AU - Flores, Tracey

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N2 - Gastrointestinal infections often induce epithelial damage that must be repaired for optimal gut function. While intestinal stem cells are critical for this regeneration process [R. C. van der Wath, B. S. Gardiner, A. W. Burgess, D. W. Smith, PLoS One 8, e73204 (2013); S. Kozar et al., Cell Stem Cell 13, 626-633 (2013)], how they are impacted by enteric infections remains poorly defined. Here, we investigate infection-mediated damage to the colonic stem cell compartment and how this affects epithelial repair and recovery from infection. Using the pathogen Clostridioides difficile, we show that infection disrupts murine intestinal cellular organization and integrity deep into the epithelium, to expose the otherwise protected stem cell compartment, in a TcdB-mediated process. Exposure and susceptibility of colonic stem cells to intoxication compromises their function during infection, which diminishes their ability to repair the injured epithelium, shown by altered stem cell signaling and a reduction in the growth of colonic organoids from stem cells isolated from infected mice. We also show, using both mouse and human colonic organoids, that TcdB from epidemic ribotype 027 strains does not require Frizzled 1/2/7 binding to elicit this dysfunctional stem cell state. This stem cell dysfunction induces a significant delay in recovery and repair of the intestinal epithelium of up to 2 wk post the infection peak. Our results uncover a mechanism by which an enteric pathogen subverts repair processes by targeting stem cells during infection and preventing epithelial regeneration, which prolongs epithelial barrier impairment and creates an environment in which disease recurrence is likely.

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Clostridioides difficile infection damages colonic stem cells via TcdB, impairing epithelial repair and recovery from disease

Abstract

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