Fecal Luminal Factors from Patients with Irritable Bowel Syndrome Induce Distinct Gene Expression of Colonoids

Overview

Journal Neurogastroenterol Motil

Specialties Gastroenterology
Neurology

Date 2022 Apr 29

PMID 35485994

Authors

Cristina Iribarren

Sofia Nordlander

Johanna Sundin

Stefan Isaksson

Otto Savolainen

Hans Tornblom

Maria K Magnusson

Magnus Simren

Lena Ohman

Affiliations

Soon will be listed here.

Abstract

Background: Alteration of the host-microbiota cross talk at the intestinal barrier may participate in the pathophysiology of irritable bowel syndrome (IBS). Therefore, we aimed to determine effects of fecal luminal factors from IBS patients on the colonic epithelium using colonoids.

Methods: Colon-derived organoid monolayers, colonoids, generated from a healthy subject, underwent stimulation with fecal supernatants from healthy subjects and IBS patients with predominant diarrhea, phosphate-buffered saline (PBS), or lipopolysaccharide (LPS). Cytokines in cell cultures and fecal LPS were measured by ELISA and mRNA gene expression of monolayers was analyzed using Qiagen RT Profiler PCR Arrays. The fecal microbiota profile was determined by the GA-map dysbiosis test and the fecal metabolite profile was analyzed by untargeted liquid chromatography/mass spectrometry.

Key Results: Colonoid monolayers stimulated with fecal supernatants from healthy subjects (n = 7), PBS (n = 4) or LPS (n = 3) presented distinct gene expression profiles, with some overlap (R Y = 0.70, Q = 0.43). Addition of fecal supernatants from healthy subjects and IBS patients (n = 9) gave rise to different gene expression profiles of the colonoid monolayers (R Y = 0.79, Q = 0.64). Genes (n = 22) related to immune response (CD1D, TLR5) and barrier integrity (CLDN15, DSC2) contributed to the separation. Levels of proinflammatory cytokines in colonoid monolayer cultures were comparable when stimulated with fecal supernatants from either donor types. Fecal microbiota and metabolite profiles, but not LPS content, differed between the study groups.

Conclusions: Fecal luminal factors from IBS patients induce a distinct colonic epithelial gene expression, potentially reflecting the disease pathophysiology. The culture of colonoids from healthy subjects with fecal supernatants from IBS patients may facilitate the exploration of IBS related intestinal micro-environmental and barrier interactions.

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