Corticosteroid enhances epithelial barrier function in intestinal organoids derived from patients with Crohn’s disease (original) (raw)
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Mechanism of glucocorticoid regulation of the intestinal tight junction barrier
American Journal of Physiology-Gastrointestinal and Liver Physiology, 2006
A defective intestinal epithelial tight junction (TJ) barrier has been proposed as an important pathogenic factor contributing to the intestinal inflammation of Crohn's disease. Glucocorticoids are first-line therapeutic agents for the treatment of moderate to severe Crohn's disease. Glucocorticoid treatment has been shown to induce retightening of the intestinal TJ barrier defect in Crohn's disease patients. However, the mechanisms that mediate the glucocorticoid therapeutic action on intestinal TJ barrier function remain unknown. The aim of this study was to elucidate the mechanism of glucocorticoid modulation of the intestinal epithelial TJ barrier using an in vitro model system. Filter-grown Caco-2 intestinal epithelial cells were used as an in vitro model to examine the effects of glucocorticoids on basal intestinal epithelial TJ barrier function and on TNF-α-induced disruption of the TJ barrier. Glucocorticoids (prednisolone and dexamethasone) did not have a signif...
PLOS ONE, 2015
Glucocorticosteroids are the first line therapy for moderate-severe flare-ups of ulcerative colitis. Despite that, up to 60% of patients do not respond adequately to steroid treatment. Previously, we reported that low IL-10 mRNA levels in intestine are associated with a poor response to glucocorticoids in active Crohn's disease. Here, we test whether IL-10 can favour the response to glucocorticoids by improving the TNFα-induced intestinal barrier damage (assessed by transepithelial electrical resistance) in Caco-2 monolayers, and their possible implications on glucocorticoid responsiveness in active ulcerative colitis. We show that the association of IL-10 and glucocorticoids improves the integrity of TNFα-treated Caco-2 cells and that p38 MAPK plays a key role. In vitro, IL-10 facilitates the nuclear translocation of p38 MAPK-phosphorylated thereby modulating glucocorticoids-receptor-α, IL-10-receptor-α and desmoglein-2 expression. In glucocorticoids-refractory patients, p38 MAPK phosphorylation and membrane desmoglein-2 expression are reduced in colonic epithelial cells. These results suggest that p38 MAPK-mediated synergism between IL-10 and glucocorticoids improves desmosome straightness contributing to the recovery of intestinal epithelium and reducing luminal antigens contact with lamina propria in ulcerative colitis. This study highlights the link between the intestinal epithelium in glucocorticoidsresponse in ulcerative colitis.
The role of the intestinal barrier function in the pathogenesis of Crohn's disease
2010
Despite of scientific efforts during the last decades, etiology and pathogenesis of the Crohn's disease (CD) remain unclear. Currently, it is widely accepted that the chronic intestinal inflammation in CD results from an inappropriate immune response that occurs in genetically susceptible individuals as the result of a complex interaction among environmental, genetic and immunological factors. A multifunctional cellular and secretory barrier separates the microbial flora from host tissues. Altered function of this barrier remains a major largely unexplored area in CD. Although there is evidence of barrier dysfunction in CD, it remains unclear whether this is a primary contributor to disease or a consequence of mucosal inflammation. The aim of this review is to summarize the recent advances in the intestinal barrier dysfunction in CD and its role in the disease pathogenesis.
Importance of disrupted intestinal barrier in inflammatory bowel diseases
Inflammatory Bowel Diseases, 2011
The current paradigm of inflammatory bowel diseases (IBD), both Crohn's disease (CD) and ulcerative colitis (UC), involves the interaction between environmental factors in the intestinal lumen and inappropriate host immune responses in genetically predisposed individuals. The intestinal mucosal barrier has evolved to maintain a delicate balance between absorbing essential nutrients while preventing the entry and responding to harmful contents. In IBD, disruptions of essential elements of the intestinal barrier lead to permeability defects. These barrier defects exacerbate the underlying immune system, subsequently resulting in tissue damage. The epithelial phenotype in active IBD is very similar in CD and UC. It is characterized by increased secretion of chloride and water, leading to diarrhea, increased permeability via both the transcellular and paracellular routes, and increased apoptosis of epithelial cells. The main cytokine that seems to drive these changes is tumor necrosis factor alpha in CD, whereas interleukin (IL)-13 may be more important in UC. Therapeutic restoration of the mucosal barrier would provide protection and prevent antigenic overload due to intestinal ''leakiness.'' Here we give an overview of the key players of the intestinal mucosal barrier and review the current literature from studies in humans and human systems on mechanisms underlying mucosal barrier dysfunction in IBD. (Inflamm Bowel Dis 2011;17:362-381)
Gastroenterology, 1998
Corticose nsitivity in¯uence s the degree and the duration of an in¯ammatory re action by alte ring targe t cell re sponse s to endoge nous and/or exoge nous glucocorticoid s. Inde e d, differe nt clinical re sponse s to glucocorticoid s have be en observe d among patie nts with Crohn' s disease , sugge sting diffe rent de gre es of corticose nsitivity in the se subje cts. The purpose of this study was to compare the corticose nsitivity of patie nts with quie scent Crohn' s dise ase to that of he althy subje cts (HS). Nine tee n patie nts with quie scent Crohn' s disease and 14 HS were studie d; all patie nts were ste roid-fre e for at least six months; 7 of the 19 were corticoste roid-de pende nt (CSD) and treate d with nonglucocortic oid immunosupp ressants at the time of the study. Corticose nsitivity was measure d by the inhibition of LPS-induce d cytokine se cretion in whole blood cell culture s tre ate d with incre asing conce ntrations (10 2 9 to 10 2 6 M) of de xame thasone . Tumor-ne crosis factor-a (TNF-a ), interle ukin-6 (IL-6), and interleukin-1b (IL-1b ) were measure d using spe ci® c immunoassays. Crohn' s dise ase patie nts had a marke dly decrease d dexamethasone -mediate d inhibition of TNF-a (P , 0.01) , IL-6 (P , 0.001) , and IL-1b (P , 0.01) compare d to he althy subje cts, with a shift of the dexame thasone dose ± response curve to the right. No signi® cant diffe rences in the basal and LPS-stimulate d secretion of the thre e cytokine s were obse rved betwe en CSD and non-CSD patie nts, and both subgroups of patie nts had similar de gre es of dexame thasone -mediate d cytokine inhibition. We conclude that patie nts with Crohn' s disease have a signi® cant decrease in the corticose nsitivity of the ir le ukocyte s. This may be re late d to a spe ci® c ge netic/constitutio nal background and/or could be acquire d, due to in¯ammation-re late d e ndocrine and/or immune factors.
Autologous organoid co-culture model reveals T cell-driven epithelial cell death in Crohn’s Disease
Frontiers in Immunology
Lympho-epithelial interactions between intestinal T resident memory cells (Trm) and the epithelium have been associated with inflammatory bowel disease (IBD) activity. We developed ex vivo autologous organoid-mucosal T cell cocultures to functionally assess lymphoepithelial interactions in Crohn’s Disease (CD) patients compared to controls. We demonstrate the direct epithelial cell death induced by autologous mucosal T cells in CD patients but not in controls. These findings were positively correlated with T cell infiltration of the organoids. This potential was inhibited by limiting lympho-epithelial interactions through CD103 and NKG2D blocking antibodies. These data directly demonstrate for the first time the direct deleterious effect of mucosal T cells on the epithelium of CD patients. Such ex-vivo models are promising techniques to unravel the pathophysiology of these diseases and the potential mode of action of current and future therapies.
Abnormal intestinal permeability in Crohn's disease pathogenesis
Annals of the New York Academy of Sciences, 2012
Increased small intestinal permeability is a longstanding observation in both Crohn's disease patients and in their healthy, asymptomatic first-degree relatives. However, the significance of this compromised gut barrier function and its place in the pathogenesis of the disease remains poorly understood. The association between abnormal small intestinal permeability and a specific mutation in the NOD2 gene, which functions to modulate both innate and adaptive immune responses to intestinal bacteria, suggests a common, genetically determined pathway by which an abnormal gut barrier could result in chronic intestinal inflammation. Furthermore, rodent colitis models show that gut barrier defects precede the development of inflammatory changes. However, it remains possible that abnormal permeability is simply a consequence of mucosal inflammation. Further insight into whether abnormal barrier function is the cause or consequence of chronic intestinal inflammation will be crucial to understanding the role of intestinal permeability in the pathogenesis of Crohn's disease.
AJP: Gastrointestinal and Liver Physiology, 2014
Barrier dysfunction is pivotal to the pathogenesis of inflammatory bowel diseases (IBD) and collagenous colitis. Glucocorticoids restore barrier function in Crohn's disease, but whether this reflects attenuated inflammation or an epithelial-specific action has not yet been addressed. Using filter-grown Caco-2 monolayers as an in vitro model of the intestinal epithelial barrier, we observed that glucocorticoids induced a time- and dose-dependent increase in transepithelial electrical resistance (TEER) in a glucocorticoid receptor-dependent manner without altering flux of larger solutes or changing principal tight junction architecture. This was accompanied by reduced paracellular cation flux, reduced expression of the pore-forming tight junction component claudin-2, and upregulation of the sealing tight junction protein claudin-4. In contrast, expression of occludin, claudin-1, -7, or -8 was not altered. Dexamethasone increased expression and activity of MAPK phosphatase-1 and in...
Characterization of Human Colon Organoids From Inflammatory Bowel Disease Patients
Frontiers in Cell and Developmental Biology, 2020
Inflammatory Bowel Diseases (IBD) are chronic inflammatory disorders, where epithelial defects drive, at least in part, some of the pathology. We reconstituted human intestinal epithelial organ, by using three-dimension culture of human colon organoids. Our aim was to characterize morphological and functional phenotypes of control (non-IBD) organoids, compared to inflamed organoids from IBD patients. The results generated describe the epithelial defects associated with IBD in primary organoid cultures, and evaluate the use of this model for pharmacological testing of anti-inflammatory approaches. Human colonic tissues were obtained from either surgical resections or biopsies, all harvested in non-inflammatory zones. Crypts were isolated from controls (non-IBD) and IBD patients and were cultured up to 12-days. Morphological (size, budding formation, polarization, luminal content), cell composition (proliferation, differentiation, immaturity markers expression), and functional (chemokine and tight junction protein expression) parameters were measured by immunohistochemistry, RT-qPCR or western-blot. The effects of inflammatory cocktail or anti-inflammatory treatments were studied in controls and IBD organoid cultures respectively. Organoid cultures from controls or IBD patients had the same cell composition after 10 to 12days of culture, but IBD organoid cultures showed an inflammatory phenotype with decreased size and budding capacity, increased cell death, luminal debris, and inverted polarization. Tight junction proteins were also significantly decreased in IBD organoid cultures. Inflammatory cytokine cocktail reproduced this inflammatory phenotype in non-IBD organoids. Clinically used treatments (5-ASA, glucocorticoids, anti-TNF) reduced some, but not all parameters. Inflammatory phenotype is associated with IBD epithelium, and can be studied in organoid cultures. This model constitutes a reliable human pre-clinical model to investigate new strategies targeting epithelial repair.