Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS

Shoko Edogawa, Adam L. Edwinson, Stephanie A. Peters, Lakshmikanth L. Chikkamenahalli, Wendy Sundt, Sara Graves, Sakteesh V. Gurunathan, Margaret Breen-Lyles, Stephen Johnson, Roy Dyer, Rondell Graham, Jun Chen, Purna Kashyap, Gianrico Farrugia, Madhusudan Grover

Research output: Contribution to journalArticle

Abstract

Objective: The intestinal lumen contains several proteases. Our aim was to determine the role of faecal proteases in mediating barrier dysfunction and symptoms in IBS. Design: 39 patients with IBS and 25 healthy volunteers completed questionnaires, assessments of in vivo permeability, ex vivo colonic barrier function in Ussing chambers, tight junction (TJ) proteins, ultrastructural morphology and 16 s sequencing of faecal microbiota rRNA. A casein-based assay was used to measure proteolytic activity (PA) in faecal supernatants (FSNs). Colonic barrier function was determined in mice (ex-germ free) humanised with microbial communities associated with different human PA states. Results: Patients with IBS had higher faecal PA than healthy volunteers. 8/20 postinfection IBS (PI-IBS) and 3/19 constipation- predominant IBS had high PA (>95th percentile). High-PA patients had more and looser bowel movements, greater symptom severity and higher in vivo and ex vivo colonic permeability. High-PA FSNs increased paracellular permeability, decreased occludin and increased phosphorylated myosin light chain (pMLC) expression. Serine but not cysteine protease inhibitor significantly blocked high-PA FSN effects on barrier. The effects on barrier were diminished by pharmacological or siRNA inhibition of protease activated receptor-2 (PAR-2). Patients with high-PA IBS had lower occludin expression, wider TJs on biopsies and reduced microbial diversity than patients with low PA. Mice humanised with high-PA IBS microbiota had greater in vivo permeability than those with low-PA microbiota. Conclusion: A subset of patients with IBS, especially in PI-IBS, has substantially high faecal PA, greater symptoms, impaired barrier and reduced microbial diversity. Commensal microbiota affects luminal PA that can influence host barrier function.

Original languageEnglish (US)
JournalGut
DOIs
StatePublished - Jan 1 2019

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Serine Proteases
Microbiota
Permeability
Occludin
Healthy Volunteers
Peptide Hydrolases
PAR-2 Receptor
Tight Junction Proteins
Cysteine Proteinase Inhibitors
Myosin Light Chains
Constipation
Caseins
Human Activities
Serine
Small Interfering RNA
Pharmacology
Biopsy

Keywords

  • Campylobacter
  • gastroenteritis
  • germ-free mice
  • microbiome
  • trypsin

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS. / Edogawa, Shoko; Edwinson, Adam L.; Peters, Stephanie A.; Chikkamenahalli, Lakshmikanth L.; Sundt, Wendy; Graves, Sara; Gurunathan, Sakteesh V.; Breen-Lyles, Margaret; Johnson, Stephen; Dyer, Roy; Graham, Rondell; Chen, Jun; Kashyap, Purna; Farrugia, Gianrico; Grover, Madhusudan.

In: Gut, 01.01.2019.

Research output: Contribution to journalArticle

Edogawa, S, Edwinson, AL, Peters, SA, Chikkamenahalli, LL, Sundt, W, Graves, S, Gurunathan, SV, Breen-Lyles, M, Johnson, S, Dyer, R, Graham, R, Chen, J, Kashyap, P, Farrugia, G & Grover, M 2019, 'Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS', Gut. https://doi.org/10.1136/gutjnl-2018-317416
Edogawa S, Edwinson AL, Peters SA, Chikkamenahalli LL, Sundt W, Graves S et al. Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS. Gut. 2019 Jan 1. https://doi.org/10.1136/gutjnl-2018-317416
Edogawa, Shoko ; Edwinson, Adam L. ; Peters, Stephanie A. ; Chikkamenahalli, Lakshmikanth L. ; Sundt, Wendy ; Graves, Sara ; Gurunathan, Sakteesh V. ; Breen-Lyles, Margaret ; Johnson, Stephen ; Dyer, Roy ; Graham, Rondell ; Chen, Jun ; Kashyap, Purna ; Farrugia, Gianrico ; Grover, Madhusudan. / Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS. In: Gut. 2019.
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abstract = "Objective: The intestinal lumen contains several proteases. Our aim was to determine the role of faecal proteases in mediating barrier dysfunction and symptoms in IBS. Design: 39 patients with IBS and 25 healthy volunteers completed questionnaires, assessments of in vivo permeability, ex vivo colonic barrier function in Ussing chambers, tight junction (TJ) proteins, ultrastructural morphology and 16 s sequencing of faecal microbiota rRNA. A casein-based assay was used to measure proteolytic activity (PA) in faecal supernatants (FSNs). Colonic barrier function was determined in mice (ex-germ free) humanised with microbial communities associated with different human PA states. Results: Patients with IBS had higher faecal PA than healthy volunteers. 8/20 postinfection IBS (PI-IBS) and 3/19 constipation- predominant IBS had high PA (>95th percentile). High-PA patients had more and looser bowel movements, greater symptom severity and higher in vivo and ex vivo colonic permeability. High-PA FSNs increased paracellular permeability, decreased occludin and increased phosphorylated myosin light chain (pMLC) expression. Serine but not cysteine protease inhibitor significantly blocked high-PA FSN effects on barrier. The effects on barrier were diminished by pharmacological or siRNA inhibition of protease activated receptor-2 (PAR-2). Patients with high-PA IBS had lower occludin expression, wider TJs on biopsies and reduced microbial diversity than patients with low PA. Mice humanised with high-PA IBS microbiota had greater in vivo permeability than those with low-PA microbiota. Conclusion: A subset of patients with IBS, especially in PI-IBS, has substantially high faecal PA, greater symptoms, impaired barrier and reduced microbial diversity. Commensal microbiota affects luminal PA that can influence host barrier function.",
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T1 - Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS

AU - Edogawa, Shoko

AU - Edwinson, Adam L.

AU - Peters, Stephanie A.

AU - Chikkamenahalli, Lakshmikanth L.

AU - Sundt, Wendy

AU - Graves, Sara

AU - Gurunathan, Sakteesh V.

AU - Breen-Lyles, Margaret

AU - Johnson, Stephen

AU - Dyer, Roy

AU - Graham, Rondell

AU - Chen, Jun

AU - Kashyap, Purna

AU - Farrugia, Gianrico

AU - Grover, Madhusudan

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Objective: The intestinal lumen contains several proteases. Our aim was to determine the role of faecal proteases in mediating barrier dysfunction and symptoms in IBS. Design: 39 patients with IBS and 25 healthy volunteers completed questionnaires, assessments of in vivo permeability, ex vivo colonic barrier function in Ussing chambers, tight junction (TJ) proteins, ultrastructural morphology and 16 s sequencing of faecal microbiota rRNA. A casein-based assay was used to measure proteolytic activity (PA) in faecal supernatants (FSNs). Colonic barrier function was determined in mice (ex-germ free) humanised with microbial communities associated with different human PA states. Results: Patients with IBS had higher faecal PA than healthy volunteers. 8/20 postinfection IBS (PI-IBS) and 3/19 constipation- predominant IBS had high PA (>95th percentile). High-PA patients had more and looser bowel movements, greater symptom severity and higher in vivo and ex vivo colonic permeability. High-PA FSNs increased paracellular permeability, decreased occludin and increased phosphorylated myosin light chain (pMLC) expression. Serine but not cysteine protease inhibitor significantly blocked high-PA FSN effects on barrier. The effects on barrier were diminished by pharmacological or siRNA inhibition of protease activated receptor-2 (PAR-2). Patients with high-PA IBS had lower occludin expression, wider TJs on biopsies and reduced microbial diversity than patients with low PA. Mice humanised with high-PA IBS microbiota had greater in vivo permeability than those with low-PA microbiota. Conclusion: A subset of patients with IBS, especially in PI-IBS, has substantially high faecal PA, greater symptoms, impaired barrier and reduced microbial diversity. Commensal microbiota affects luminal PA that can influence host barrier function.

AB - Objective: The intestinal lumen contains several proteases. Our aim was to determine the role of faecal proteases in mediating barrier dysfunction and symptoms in IBS. Design: 39 patients with IBS and 25 healthy volunteers completed questionnaires, assessments of in vivo permeability, ex vivo colonic barrier function in Ussing chambers, tight junction (TJ) proteins, ultrastructural morphology and 16 s sequencing of faecal microbiota rRNA. A casein-based assay was used to measure proteolytic activity (PA) in faecal supernatants (FSNs). Colonic barrier function was determined in mice (ex-germ free) humanised with microbial communities associated with different human PA states. Results: Patients with IBS had higher faecal PA than healthy volunteers. 8/20 postinfection IBS (PI-IBS) and 3/19 constipation- predominant IBS had high PA (>95th percentile). High-PA patients had more and looser bowel movements, greater symptom severity and higher in vivo and ex vivo colonic permeability. High-PA FSNs increased paracellular permeability, decreased occludin and increased phosphorylated myosin light chain (pMLC) expression. Serine but not cysteine protease inhibitor significantly blocked high-PA FSN effects on barrier. The effects on barrier were diminished by pharmacological or siRNA inhibition of protease activated receptor-2 (PAR-2). Patients with high-PA IBS had lower occludin expression, wider TJs on biopsies and reduced microbial diversity than patients with low PA. Mice humanised with high-PA IBS microbiota had greater in vivo permeability than those with low-PA microbiota. Conclusion: A subset of patients with IBS, especially in PI-IBS, has substantially high faecal PA, greater symptoms, impaired barrier and reduced microbial diversity. Commensal microbiota affects luminal PA that can influence host barrier function.

KW - Campylobacter

KW - gastroenteritis

KW - germ-free mice

KW - microbiome

KW - trypsin

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