Neutrophils prevent rectal bleeding in ulcerative colitis by peptidyl-arginine deiminase-4-dependent immunothrombosis

Leppkes M, Lindemann A, Goesswein S, Paulus S, Roth D, Hartung A, Liebing E, Zundler S, Gonzalez-Acera M, Patankar J, Mascia F, Scheibe K, Hoffmann M, Uderhardt S, Schauer C, Foersch S, Neufert C, Vieth M, Schett G, Atreya R, Kuehl AA, Bleich A, Becker C, Herrmann M, Neurath M (2021)

Publication Type: Journal article

Publication year: 2021

Journal

Gut BMJ Publishing Group

DOI: 10.1136/gutjnl-2021-324725

Abstract

Objective Bleeding ulcers and erosions are hallmarks of active ulcerative colitis (UC). However, the mechanisms controlling bleeding and mucosal haemostasis remain elusive. Design We used high-resolution endoscopy and colon tissue samples of active UC (n = 36) as well as experimental models of physical and chemical mucosal damage in mice deficient for peptidyl-arginine deiminase-4 (PAD4), gnotobiotic mice and controls. We employed endoscopy, histochemistry, live-cell microscopy and flow cytometry to study eroded mucosal surfaces during mucosal haemostasis. Results Erosions and ulcerations in UC were covered by fresh blood, haematin or fibrin visible by endoscopy. Fibrin layers rather than fresh blood or haematin on erosions were inversely correlated with rectal bleeding in UC. Fibrin layers contained ample amounts of neutrophils coaggregated with neutrophil extracellular traps (NETs) with detectable activity of PAD. Transcriptome analyses showed significantly elevated PAD4 expression in active UC. In experimentally inflicted wounds, we found that neutrophils underwent NET formation in a PAD4-dependent manner hours after formation of primary blood clots, and remodelled clots to immunothrombi containing citrullinated histones, even in the absence of microbiota. PAD4-deficient mice experienced an exacerbated course of dextrane sodium sulfate-induced colitis with markedly increased rectal bleeding (96 % vs 10 %) as compared with controls. PAD4-deficient mice failed to remodel blood clots on mucosal wounds eliciting impaired healing. Thus, NET-associated immunothrombi are protective in acute colitis, while insufficient immunothrombosis is associated with rectal bleeding. Conclusion Our findings uncover that neutrophils induce secondary immunothrombosis by PAD4-dependent mechanisms. Insufficient immunothrombosis may favour rectal bleeding in UC.

Authors with CRIS profile

Moritz Leppkes Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Aylin Lindemann Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Susanne Paulus Lehrstuhl für Innere Medizin I Dominik Roth Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Eva Liebing Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Sebastian Zundler Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Jay Patankar Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Fabrizio Mascia Medizinische Fakultät Kristina Koop Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology Markus Hoffmann Department of Medicine 3 – Rheumatology and Immunology Stefan Uderhardt Juniorprofessur für Rheumatologie (Stiftungsprofessur) Christine Schauer Department of Medicine 3 – Rheumatology and Immunology Clemens Neufert Medizinische Fakultät Michael Vieth Medizinische Fakultät Georg Schett Lehrstuhl für Innere Medizin III Raja Atreya Professur für Translationale Immunforschung bei chronisch entzündlichen Darmerkrankungen (Heisenberg-Professur) Christoph Becker Professur für Molekulare Gastroenterologie Martin Herrmann Department of Medicine 3 – Rheumatology and Immunology Markus Neurath Lehrstuhl für Innere Medizin I

Additional Organisation(s)

Interdisziplinäres Zentrum für Klinische Forschung

Related research project(s)

J085: Analysis of cell-type-specific roles and crosstalk of IL36R signaling in gut inflammation and fibrosis Nov. 16, 2020 - Nov. 15, 2023

Involved external institutions

Paracelsus-Elena-Klinik Kassel DE Germany (DE) Medizinische Hochschule Hannover (MHH) / Hannover Medical School DE Germany (DE) Charité - Universitätsmedizin Berlin DE Germany (DE)

How to cite

APA:

Leppkes, M., Lindemann, A., Goesswein, S., Paulus, S., Roth, D., Hartung, A.,... Neurath, M. (2021). Neutrophils prevent rectal bleeding in ulcerative colitis by peptidyl-arginine deiminase-4-dependent immunothrombosis. Gut. https://dx.doi.org/10.1136/gutjnl-2021-324725

MLA:

Leppkes, Moritz, et al. "Neutrophils prevent rectal bleeding in ulcerative colitis by peptidyl-arginine deiminase-4-dependent immunothrombosis." Gut (2021).

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