Fecal luminal factors from patients with irritable bowel syndrome induce distinct gene expression of colonoids
Artikel i vetenskaplig tidskrift, 2022

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 RT2 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 (R2Y = 0.70, Q2= 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 (R2Y = 0.79, Q2= 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.

epithelial barrier

colonoids

irritable bowel syndrome

host-microbiota cross talk

intestinal microenvironment

Författare

Cristina Iribarren

Göteborgs universitet

Sofia Nordlander

Göteborgs universitet

Johanna Sundin

Göteborgs universitet

Stefan Isaksson

Göteborgs universitet

Otto Savolainen

Chalmers, Biologi och bioteknik, Systembiologi, CMSI

H. Törnblom

Göteborgs universitet

Maria K. Magnusson

Göteborgs universitet

M. Simren

The University of North Carolina at Chapel Hill

Göteborgs universitet

Lena Öhman

Göteborgs universitet

Neurogastroenterology and Motility

1350-1925 (ISSN)

Vol. In Press

Ämneskategorier

Immunologi

Gastroenterologi

Neurologi

DOI

10.1111/nmo.14390

Mer information

Senast uppdaterat

2022-05-11