Establishment and validation of organ-on-chip models for Inflammatory Bowel Disease modelling in vitro

Inflammatory Bowel Disease is a chronic disorder resulting in inflammation and ulceration of the intestinal mucosa. It is a multifactorial disease caused by the interplay between genetics of an individual, microbiota and environment. A number of genes have been reported as susceptibility factors, including NOD2 that plays a role in the recognition of bacterial cell wall components and activation of pro-inflammatory pathways. The main aspects of the disease are disruption of the intestinal barrier and upregulation in the expression of pro-inflammatory cytokines. Standard 2D models of the intestine fail to mimic physiological complexity of the intestinal tissue inclusive of presence of various cell types, extracelular matrix and incorporation of the fluid flow. There is a need for novel in vitro models that would mimic human biology more closely while being suitable for including in the drug development pipeline. In this thesis we present the development of multiple gut on a chip models that capture boundary aspect of the intestinal tissue. We investagated epithelial immune response to the pro-inflammatory triggers characteristic to IBD in mono- and co-culture with hematopoietic cells. We decribe generation of the models established with commonly used Caco-2 adenocarcinoma cell line, iPSC-derived as well as human stem cells derived intestinal organoids. Our results show that organ on a chip technology could be successfully used to model human intestine. OrganoPlate® platform used in the development of the models suited for high throughput screening allowed for generation of physiologically relevant models with pump-free perfusion. Obtained epithelial barriers exhibited rapid cell polarization, expression of major intestinal markers and barrier integrity. We show a proof of concept study in which iPSC-derived intestinal organoids cultured in a tubular form were triggered with pro-inflammatory cytokines. The inflamed phenotype was efficiently reverted with the addition of an anti-inflammatory compound proving suitability of the model for IBD-related drug discovery research. These findings provide the basis for further research towards generation of novel in vitro models. Combining organoid cultures with organ on chip technology advances the current state of the art in tissue and disease modelling giving a promise for significant developments in the drug discovery field.