An autologous colonic organoid‐derived monolayer model to study immune: bacterial interactions in Crohn's disease patients

Abstract

Objectives Crohn's disease (CD) initiation and pathogenesis are believed to involve an environmental trigger in a genetically susceptible person that results in an immune response against commensal gut bacteria, leading to a compromised intestinal epithelial barrier and a cycle of inflammation. However, it has been difficult to study the contribution of all factors together in a physiologically relevant model and in a heterogenous patient population. Methods We developed an autologous colonic monolayer model that incorporated the immune response from the same donor and a commensal bacteria, Faecalibacterium prausnitzii . Two‐dimensional monolayers were grown from three‐dimensional organoids generated from intestinal biopsies, and the epithelial integrity of the epithelium was measured using transepithelial electrical resistance. We determined the effect of immune cells alone, bacteria alone and the co‐culture of immune cells and bacteria on integrity. Results Monolayers derived from CD donors had impaired epithelial integrity compared to those from non‐inflammatory bowel disease (IBD) donors. This integrity was further impaired by culture with bacteria, but not immune cells, despite a higher frequency of inflammatory phenotype peripheral T cells in CD donors. Variability in epithelial integrity was higher in CD donors than in non‐IBD donors. Conclusion We have developed a new autologous model to study the complexity of CD, which allows for the comparison of the barrier properties of the colonic epithelium and the ability to study how autologous immune cells directly affect the colonic barrier and whether this is modified by luminal bacteria. This new model allows for the study of individual patients and could inform treatment decisions.

Molecular and Functional Characterization of Human Intestinal Organoids and Monolayers for Modeling Epithelial Barrier

“…Recent studies suggest IBD-PDOs and non-IBD-PDOs have distinct phenotypes with respect to transcriptional signatures 39 and baseline permeability. 40 However, transcriptional variation in our study was more strongly associated with growth media, culture conditions, and donor variability than with disease status ( Figure 3A ). Moreover, control- and IBD-PDOs consistently formed monolayers with stable TEER that were impermeable to 0.4 kDa Lucifer yellow ( Figure 5 and S5 ).…”

The Application of Intestinal Stem Cell Organoids in the Treatment of Inflammatory Bowel Disease

“…Testing of current medications on patient-specific organoids will optimize treatment for patients, reduce therapy failure and mitigate adverse effects [15]. Organoids can be used to produce colonic monolayers where the individual effects of immune cells and bacteria can be assessed in CD patients [24]. Additionally, organoids offer a new https://doi.org/10.46889/JRMBR.2024.5102 https://athenaeumpub.com/journal-of-regenerative-medicine-biology-research/ therapeutic approach to reconstruct the epithelial barrier in the inflamed mucosa of IBD patients to accelerate healing through engraftment onto damaged epithelium [16,19].…”

Human intestinal organoids: Modeling gastrointestinal physiology and immunopathology — current applications and limitations

“…T cells are known to drive the pathogenesis of intestinal inflammatory disorders due to excessive cytotoxicity and production of pro-inflammatory cytokines. T cells and organoids have been co-cultured indirectly using transwell systems (with T cells cultured in the basolateral chamber) [87], or directly by embedding both populations in an ECM [88,89]. Using the indirect coculture method, it was demonstrated that secretion of inflammatory mediators by T cells is sufficient to destroy colonoid monolayers [87].…”

“…T cells and organoids have been co-cultured indirectly using transwell systems (with T cells cultured in the basolateral chamber) [87], or directly by embedding both populations in an ECM [88,89]. Using the indirect coculture method, it was demonstrated that secretion of inflammatory mediators by T cells is sufficient to destroy colonoid monolayers [87]. An autologous co-culture system (cell populations obtained from the same patient) was developed to study disease and patient-specific interactions between mucosal lymphocytes and enteroids [88].…”