Biopsy-derived Intestinal Epithelial Cell Cultures for Pathway-based Stratification of Patients With Inflammatory Bowel Disease

Vanhove, Wiebe; Nys, Kris; Arijs, Ingrid; Cleynen, Isabelle; Noben, Manuel; Schepper, Sebastiaan De; Assche, Gert Van; Ferrante, Marc; Vermeire, Séverine

doi:10.1093/ecco-jcc/jjx122

Cited by 14 publications

(20 citation statements)

References 52 publications

(60 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intestinal crypts were isolated from 4–6 colon biopsies per individual as described before. 27 At the end of the procedure, the crypts were re-suspended in Matrigel [Growth Factor Reduced, phenol-red-free, Corning, NY, USA] diluted with basal medium [50/50%]. 28 To allow the formation of 3D organoid structures, three droplets [13.3 μl] of each cell suspension were plated in every well of a 24-well tissue culture plate [8–12 wells/4–6 biopsies dependent on biopsy size and isolation efficiency].…”

Section: Methodsmentioning

confidence: 99%

Butyrate Does Not Protect Against Inflammation-induced Loss of Epithelial Barrier Function and Cytokine Production in Primary Cell Monolayers From Patients With Ulcerative Colitis

Vancamelbeke

Laeremans

Vanhove

et al. 2019

Journal of Crohn's and Colitis

Self Cite

View full text Add to dashboard Cite

Background and Aims In vitro studies using immortalised cancer cell lines showed that butyrate has an overall positive effect on epithelial barrier integrity, but the physiological relevance of cancer cell lines is limited. We developed epithelial monolayers from human tissue samples of patients with ulcerative colitis [UC] to assess the effect of butyrate on epithelial barrier function. Methods A protocol to establish monolayers from primary epithelial cells of UC patients [n = 10] and non-UC controls [n = 10] was optimised. The monolayers were treated with 8 mM sodium butyrate ± tumour necrosis factor alpha [TNFα] and type II interferon [IFNγ] for 48 h. Changes in transepithelial electrical resistance were monitored. Barrier gene expression levels were measured. Inflammatory proteins in the supernatant of the cells were quantified with OLINK. Results We demonstrated that primary monolayer cultures can be grown within 1 week of culture with robust resistance values and polarised tight junction expression. Butyrate treatment of the cultures increased resistance but was detrimental in combination with TNFα and IFNγ. The combined treatment further induced even higher IL8 mRNA and inflammatory protein secretion than for the inflammatory mediators alone. The observed effects were similar in cultures from patients and non-UC controls, suggesting that there were no patient-specific responses responsible for these findings. Conclusions We found that butyrate does not protect against inflammation-induced barrier dysfunction and even worsens its effects in primary epithelial monolayers of UC patients and controls. The basic mechanisms of butyrate should therefore be reconsidered in future studies, in particular in patients with active inflammation and pre-existing barrier defects as is known for UC.

show abstract

Section: Methodsmentioning

confidence: 99%

Butyrate Does Not Protect Against Inflammation-induced Loss of Epithelial Barrier Function and Cytokine Production in Primary Cell Monolayers From Patients With Ulcerative Colitis

Vancamelbeke

Laeremans

Vanhove

et al. 2019

Journal of Crohn's and Colitis

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, mice deficient in both Atg16l1 and Xbp1 (X-box binding protein 1) specifically in IECs ( [45], which seems to be due to defective ERN1 degradation by autophagy [44]. Primary IECs from CD patients carrying several risk variants implicated in both autophagy and ER stress, but not in autophagy alone, exhibit significantly higher ER stress response after ER stress induction but not at the basal level, indicating that the genetic predispositions in both pathways should be considered when looking at functional level [48].…”

Section: Autophagy and Er Stressmentioning

confidence: 99%

New insights into the interplay between autophagy, gut microbiota and inflammatory responses in IBD

2019

View full text Add to dashboard Cite

One of the most significant challenges of inflammatory bowel disease (IBD) research is to understand how alterations in the symbiotic relationship between the genetic composition of the host and the intestinal microbiota, under impact of specific environmental factors, lead to chronic intestinal inflammation. Genome-wide association studies, followed by functional studies, have identified a role for numerous autophagy genes in IBD, especially in Crohn disease. Studies using in vitro and in vivo models, in addition to human clinical studies have revealed that autophagy is pivotal for intestinal homeostasis maintenance, gut ecology regulation, appropriate intestinal immune responses and anti-microbial protection. This review describes the latest researches on the mechanisms by which dysfunctional autophagy leads to disrupted intestinal epithelial function, gut dysbiosis, defect in anti-microbial peptide secretion by Paneth cells, endoplasmic reticulum stress response and aberrant immune responses to pathogenic bacteria. A better understanding of the role of autophagy in IBD pathogenesis may provide better sub-classification of IBD phenotypes and novel approaches for disease management.

show abstract

“…Organoids can be established from a single ISC in vitro [ 15 ], and faithfully retain the physiological and pathological features of their tissue of origin [ 16 , 17 ]. Thus, organoids have been used to dissect underlying pathologic changes in various gastrointestinal disease [ 13 , 18 – 20 ], in addition to the 2D-culture system [ 21 ]. A recent study identified permanent changes in gene expression patterns of colonic organoids established from ulcerative colitis (UC) patients [ 22 ], and indicated that colonic ISCs carry imprinted alterations possibly contributing to the perpetuation of the disease.…”

Section: Introductionmentioning

confidence: 99%

Single cell analysis of Crohn’s disease patient-derived small intestinal organoids reveals disease activity-dependent modification of stem cell properties

et al. 2018

View full text Add to dashboard Cite

BackgroundIntestinal stem cells (ISCs) play indispensable roles in the maintenance of homeostasis, and also in the regeneration of the damaged intestinal epithelia. However, whether the inflammatory environment of Crohn’s disease (CD) affects properties of resident small intestinal stem cells remain uncertain.MethodsCD patient-derived small intestinal organoids were established from enteroscopic biopsy specimens taken from active lesions (aCD-SIO), or from mucosa under remission (rCD-SIO). Expression of ISC-marker genes in those organoids was examined by immunohistochemistry, and also by microfluid-based single-cell multiplex gene expression analysis. The ISC-specific function of organoid cells was evaluated using a single-cell organoid reformation assay.ResultsISC-marker genes, OLFM4 and SLC12A2, were expressed by an increased number of small intestinal epithelial cells in the active lesion of CD. aCD-SIOs, rCD-SIOs or those of non-IBD controls (NI-SIOs) were successfully established from 9 patients. Immunohistochemistry showed a comparable level of OLFM4 and SLC12A2 expression in all organoids. Single-cell gene expression data of 12 ISC-markers were acquired from a total of 1215 cells. t-distributed stochastic neighbor embedding analysis identified clusters of candidate ISCs, and also revealed a distinct expression pattern of SMOC2 and LGR5 in ISC-cluster classified cells derived from aCD-SIOs. Single-cell organoid reformation assays showed significantly higher reformation efficiency by the cells of the aCD-SIOs compared with that of cells from NI-SIOs.ConclusionsaCD-SIOs harbor ISCs with modified marker expression profiles, and also with high organoid reformation ability. Results suggest modification of small intestinal stem cell properties by unidentified factors in the inflammatory environment of CD.Electronic supplementary materialThe online version of this article (10.1007/s00535-018-1437-3) contains supplementary material, which is available to authorized users.

show abstract

Biopsy-derived Intestinal Epithelial Cell Cultures for Pathway-based Stratification of Patients With Inflammatory Bowel Disease

Cited by 14 publications

References 52 publications

Butyrate Does Not Protect Against Inflammation-induced Loss of Epithelial Barrier Function and Cytokine Production in Primary Cell Monolayers From Patients With Ulcerative Colitis

Butyrate Does Not Protect Against Inflammation-induced Loss of Epithelial Barrier Function and Cytokine Production in Primary Cell Monolayers From Patients With Ulcerative Colitis

New insights into the interplay between autophagy, gut microbiota and inflammatory responses in IBD

Single cell analysis of Crohn’s disease patient-derived small intestinal organoids reveals disease activity-dependent modification of stem cell properties

Contact Info

Product

Resources

About