Major Toxins Remodel the Intestinal Epithelia, Affecting Spore Adherence/internalization into Intestinal Tissue and Their Association with Gut Vitronectin

Overview

Journal bioRxiv

Date 2025 Feb 20

PMID 39974910

Authors

Pablo Castro-Cordova

Osiris K Lopez-Garcia

Josue Orozco

Nicolas Montes-Bravo

Fernando Gil

Marjorie Pizarro-Guajardo

Daniel Paredes-Sabja

Affiliations

Soon will be listed here.

Abstract

The most common cause of healthcare-associated diarrhea and colitis in the U.S., is , a spore-forming pathogen. Two toxins, TcdA and TcdB, are major virulence factors essential for disease manifestations, while spores are essential for disease transmission and recurrence. Both toxins cause major damage to the epithelial barrier, trigger massive inflammation, and reshape the microbiome and metabolic composition, facilitating colonization. spores, essential for transmission and recurrence of the disease, persist adhered and internalized in the intestinal epithelia. Studies have suggested that toxin-neutralization in combination with antibiotic during CDI treatment in humans significantly reduces disease recurrence, suggesting a link between toxin-mediated damage and spore persistence. Here, we show that TcdA/TcdB-intoxication of intestinal epithelial Caco-2 cells leads to remodeling of accessible levels of fibronectin (Fn) and vitronectin (Vn) and their cognate alpha-integrin subunits. While TcdB-intoxication of intestinal tissue had no impact in accessible levels of Fn and Vn, but significantly increased levels of intracellular Vn. We observed that Fn and Vn released to the supernatant readily bind to spores , while TcdB-intoxication of intestinal tissue led to increased association of spores with gut Vn. Toxin-intoxication of the intestinal tissue also contributes to increased adherence and internalization of spores. However, TcdB-intoxicated ligated loops infected of mice treated with Bezlotoxumanb (monoclonal anti-TcdB antibodies) did not prevent TcdB-mediated increased spore adherence and internalization into intestinal tissue. This study highlights the importance of studying the impact of toxins of host tissues has in interaction with host surfaces that may contribute to increased persistence and disease recurrence.

References

Peters J, Loredo G, Chen G, Maunder R, Hahn T, Willits N . Plasma levels of fibronectin bearing the alternatively spliced EIIIB segment are increased after major trauma. J Lab Clin Med. 2003; 141(6):401-10. DOI: 10.1016/S0022-2143(03)00042-8. View

LaFrance M, Farrow M, Chandrasekaran R, Sheng J, Rubin D, Lacy D . Identification of an epithelial cell receptor responsible for Clostridium difficile TcdB-induced cytotoxicity. Proc Natl Acad Sci U S A. 2015; 112(22):7073-8. PMC: 4460460. DOI: 10.1073/pnas.1500791112. View

Chumbler N, Farrow M, Lapierre L, Franklin J, Haslam D, Haslam D . Clostridium difficile Toxin B causes epithelial cell necrosis through an autoprocessing-independent mechanism. PLoS Pathog. 2012; 8(12):e1003072. PMC: 3516567. DOI: 10.1371/journal.ppat.1003072. View

Abdullayeva G, Liu H, Liu T, Simmons A, Novelli M, Huseynova I . Goblet cell differentiation subgroups in colorectal cancer. Proc Natl Acad Sci U S A. 2024; 121(43):e2414213121. PMC: 11513979. DOI: 10.1073/pnas.2414213121. View

Martens E, Neumann M, Desai M . Interactions of commensal and pathogenic microorganisms with the intestinal mucosal barrier. Nat Rev Microbiol. 2018; 16(8):457-470. DOI: 10.1038/s41579-018-0036-x. View

Schnizlein M, Young V . Capturing the environment of the Clostridioides difficile infection cycle. Nat Rev Gastroenterol Hepatol. 2022; 19(8):508-520. DOI: 10.1038/s41575-022-00610-0. View

Tao L, Zhang J, Meraner P, Tovaglieri A, Wu X, Gerhard R . Frizzled proteins are colonic epithelial receptors for C. difficile toxin B. Nature. 2016; 538(7625):350-355. PMC: 5519134. DOI: 10.1038/nature19799. View

Castellanos M, Leira R, Serena J, Blanco M, Pedraza S, Castillo J . Plasma cellular-fibronectin concentration predicts hemorrhagic transformation after thrombolytic therapy in acute ischemic stroke. Stroke. 2004; 35(7):1671-6. DOI: 10.1161/01.STR.0000131656.47979.39. View

Chaves-Olarte E, Weidmann M, Thelestam M . Toxins A and B from Clostridium difficile differ with respect to enzymatic potencies, cellular substrate specificities, and surface binding to cultured cells. J Clin Invest. 1997; 100(7):1734-41. PMC: 508356. DOI: 10.1172/JCI119698. View

10.

Nikitas G, Deschamps C, Disson O, Niault T, Cossart P, Lecuit M . Transcytosis of Listeria monocytogenes across the intestinal barrier upon specific targeting of goblet cell accessible E-cadherin. J Exp Med. 2011; 208(11):2263-77. PMC: 3201198. DOI: 10.1084/jem.20110560. View

11.

Henderson B, Nair S, Pallas J, Williams M . Fibronectin: a multidomain host adhesin targeted by bacterial fibronectin-binding proteins. FEMS Microbiol Rev. 2010; 35(1):147-200. DOI: 10.1111/j.1574-6976.2010.00243.x. View

12.

Warn P, Thommes P, Sattar A, Corbett D, Flattery A, Zhang Z . Disease Progression and Resolution in Rodent Models of Clostridium difficile Infection and Impact of Antitoxin Antibodies and Vancomycin. Antimicrob Agents Chemother. 2016; 60(11):6471-6482. PMC: 5075051. DOI: 10.1128/AAC.00974-16. View

13.

Lee J, Kim H, Cha M, Park H, Kim Y, Kim I . Clostridium difficile toxin A promotes dendritic cell maturation and chemokine CXCL2 expression through p38, IKK, and the NF-kappaB signaling pathway. J Mol Med (Berl). 2008; 87(2):169-80. DOI: 10.1007/s00109-008-0415-2. View

14.

Wohlan K, Goy S, Olling A, Srivaratharajan S, Tatge H, Genth H . Pyknotic cell death induced by Clostridium difficile TcdB: chromatin condensation and nuclear blister are induced independently of the glucosyltransferase activity. Cell Microbiol. 2014; 16(11):1678-92. DOI: 10.1111/cmi.12317. View

15.

Vachon P, Simoneau A, Beaulieu J . Cellular fibronectin expression is down-regulated at the mRNA level in differentiating human intestinal epithelial cells. Exp Cell Res. 1995; 216(1):30-4. DOI: 10.1006/excr.1995.1004. View

16.

Farrow M, Chumbler N, Lapierre L, Franklin J, Rutherford S, Goldenring J . Clostridium difficile toxin B-induced necrosis is mediated by the host epithelial cell NADPH oxidase complex. Proc Natl Acad Sci U S A. 2013; 110(46):18674-9. PMC: 3831945. DOI: 10.1073/pnas.1313658110. View

17.

Giuffrida P, Pinzani M, Corazza G, Di Sabatino A . Biomarkers of intestinal fibrosis - one step towards clinical trials for stricturing inflammatory bowel disease. United European Gastroenterol J. 2016; 4(4):523-30. PMC: 4971797. DOI: 10.1177/2050640616640160. View

18.

Peritore-Galve F, Shupe J, Cave R, Childress K, Washington M, Kuehne S . Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. PLoS Pathog. 2022; 18(2):e1010323. PMC: 8890742. DOI: 10.1371/journal.ppat.1010323. View

19.

Yuan P, Zhang H, Cai C, Zhu S, Zhou Y, Yang X . Chondroitin sulfate proteoglycan 4 functions as the cellular receptor for Clostridium difficile toxin B. Cell Res. 2014; 25(2):157-68. PMC: 4650570. DOI: 10.1038/cr.2014.169. View

20.

Podack E, Preissner K, MULLER-EBERHARD H . Inhibition of C9 polymerization within the SC5b-9 complex of complement by S-protein. Acta Pathol Microbiol Immunol Scand Suppl. 1984; 284:89-96. View