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A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins

Background: Clostridioides difficile toxins TcdA and TcdB are responsible for diarrhea and colitis. Lack of functional studies in organoid models of the gut prompted us to elucidate the toxin’s effects on epithelial barrier function and the molecular mechanisms for diarrhea and inflammation. Methods...

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Autores principales: Schneemann, Martina, Heils, Lucas, Moos, Verena, Weiß, Franziska, Krug, Susanne M., Weiner, January, Beule, Dieter, Gerhard, Ralf, Schulzke, Jörg-Dieter, Bücker, Roland
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10674794/
https://www.ncbi.nlm.nih.gov/pubmed/37999506
http://dx.doi.org/10.3390/toxins15110643
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author Schneemann, Martina
Heils, Lucas
Moos, Verena
Weiß, Franziska
Krug, Susanne M.
Weiner, January
Beule, Dieter
Gerhard, Ralf
Schulzke, Jörg-Dieter
Bücker, Roland
author_facet Schneemann, Martina
Heils, Lucas
Moos, Verena
Weiß, Franziska
Krug, Susanne M.
Weiner, January
Beule, Dieter
Gerhard, Ralf
Schulzke, Jörg-Dieter
Bücker, Roland
author_sort Schneemann, Martina
collection PubMed
description Background: Clostridioides difficile toxins TcdA and TcdB are responsible for diarrhea and colitis. Lack of functional studies in organoid models of the gut prompted us to elucidate the toxin’s effects on epithelial barrier function and the molecular mechanisms for diarrhea and inflammation. Methods: Human adult colon organoids were cultured on membrane inserts. Tight junction (TJ) proteins and actin cytoskeleton were analyzed for expression via Western blotting and via confocal laser-scanning microscopy for subcellular localization. Results: Polarized intestinal organoid monolayers were established from stem cell-containing colon organoids to apply toxins from the apical side and to perform functional measurements in the organoid model. The toxins caused a reduction in transepithelial electrical resistance in human colonic organoid monolayers with sublethal concentrations. Concomitantly, we detected increased paracellular permeability fluorescein and FITC-dextran-4000. Human colonic organoid monolayers exposed to the toxins exhibited redistribution of barrier-forming TJ proteins claudin-1, -4 and tricellulin, whereas channel-forming claudin-2 expression was increased. Perijunctional F-actin cytoskeleton organization was affected. Conclusions: Adult stem cell-derived human colonic organoid monolayers were applicable as a colon infection model for electrophysiological measurements. The TJ changes noted can explain the epithelial barrier dysfunction and diarrhea in patients, as well as increased entry of luminal antigens triggering inflammation.
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spelling pubmed-106747942023-11-04 A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins Schneemann, Martina Heils, Lucas Moos, Verena Weiß, Franziska Krug, Susanne M. Weiner, January Beule, Dieter Gerhard, Ralf Schulzke, Jörg-Dieter Bücker, Roland Toxins (Basel) Article Background: Clostridioides difficile toxins TcdA and TcdB are responsible for diarrhea and colitis. Lack of functional studies in organoid models of the gut prompted us to elucidate the toxin’s effects on epithelial barrier function and the molecular mechanisms for diarrhea and inflammation. Methods: Human adult colon organoids were cultured on membrane inserts. Tight junction (TJ) proteins and actin cytoskeleton were analyzed for expression via Western blotting and via confocal laser-scanning microscopy for subcellular localization. Results: Polarized intestinal organoid monolayers were established from stem cell-containing colon organoids to apply toxins from the apical side and to perform functional measurements in the organoid model. The toxins caused a reduction in transepithelial electrical resistance in human colonic organoid monolayers with sublethal concentrations. Concomitantly, we detected increased paracellular permeability fluorescein and FITC-dextran-4000. Human colonic organoid monolayers exposed to the toxins exhibited redistribution of barrier-forming TJ proteins claudin-1, -4 and tricellulin, whereas channel-forming claudin-2 expression was increased. Perijunctional F-actin cytoskeleton organization was affected. Conclusions: Adult stem cell-derived human colonic organoid monolayers were applicable as a colon infection model for electrophysiological measurements. The TJ changes noted can explain the epithelial barrier dysfunction and diarrhea in patients, as well as increased entry of luminal antigens triggering inflammation. MDPI 2023-11-04 /pmc/articles/PMC10674794/ /pubmed/37999506 http://dx.doi.org/10.3390/toxins15110643 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Schneemann, Martina
Heils, Lucas
Moos, Verena
Weiß, Franziska
Krug, Susanne M.
Weiner, January
Beule, Dieter
Gerhard, Ralf
Schulzke, Jörg-Dieter
Bücker, Roland
A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title_full A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title_fullStr A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title_full_unstemmed A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title_short A Colonic Organoid Model Challenged with the Large Toxins of Clostridioides difficile TcdA and TcdB Exhibit Deregulated Tight Junction Proteins
title_sort colonic organoid model challenged with the large toxins of clostridioides difficile tcda and tcdb exhibit deregulated tight junction proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10674794/
https://www.ncbi.nlm.nih.gov/pubmed/37999506
http://dx.doi.org/10.3390/toxins15110643
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