A Micro-engineered Human Colon Intestine-Chip Platform to Study Leaky Barrier

Abstract

The intestinal epithelial barrier supports the symbiotic relationship between the microbiota colonizing the intestinal epithelium and the host immune system to maintain homeostasis. Leaky barrier is increasingly recognized as part of the pathogenesis of a number of chronic conditions in addition to inflammatory and infectious diseases. As our understanding on the regulation of the barrier remains limited, effective therapeutic targeting for the compromised barrier is still an unmet need. Here we combined advancements on the organoids and Organ-on-Chip technologies to establish a micro-engineered Colon Intestine-Chip for studying development and regulation of the human intestinal barrier. Our data demonstrate the significance of the endothelium in co-culture with the epithelial cells within a tissue-relevant microenvironment for the establishment of a tight epithelial barrier of polarized cells. Pathway analysis of the RNA sequencing (RNA-Seq), revealed significant upregulation of mechanisms relevant to the maturation of the intestinal epithelium in organoid-derived epithelial cells in co-culture with endothelium as compared to organoids maintained in suspension. We provide evidence that the Colon Intestine-Chip platform responds to interferon gamma (IFNγ), a prototype cytokine utilized to model inflammation-induced barrier disruption, by induction of apoptosis and reorganization of the apical junctional complexes as shown with other systems. We also describe the mechanism of action of interleukin 22 (IL-22) on mature, organoid-derived intestinal epithelial cells that is consistent with barrier disruption. Overall we propose the Colon Intestine-Chip as a promising human organoid-derived platform to decipher mechanisms driving the development of leaky gut in patients and enable their translation for this unmet medical need.

Effects of Human Milk Oligosaccharides on the Adult Gut Microbiota and Barrier Function

“…To establish Colon Intestine-Chips, colonic organoids were seeded on S-1 chips (Emulate Inc., Boston, MA, USA) and processed as follows [36](Apostolou et al, unpublished): Chips were washed with 70% ethanol, cell culture water, and ER2 reagent (Emulate Inc.), and then activated with ER1 (Emulate Inc.) working solution (0.5 mg/mL ER1 powder in ER2 solution) using 20 min UV light treatment (Nailstar, NS-01 EU/UK). The chips were, then, washed with ER2 and PBS and the inner surfaces of both channels and the porous PDMS membrane coated with extracellular matrix proteins (ECM).…”

Human immunocompetent Organ-on-Chip platforms allow safety profiling of tumor-targeted T-cell bispecific antibodies

“…S6 and S7A), where they form a tight, polarized barrier, containing the full range of mature intestinal cell types. We have previously shown that the inclusion of luminal flow, peristaltic motion, and an endothelial layer enhances the maturation and physiological fidelity of the barrier, compared with organoids in conventional 3D culture(35).…”

Gastrointestinal organs and organoids-on-a-chip: advances and translation into the clinics

“…Different models of gastrointestinal organs and organoids-on-a-chip have been innovatively explored in drug testing, some of which are already GOoC models, summarized in table 1 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44].…”

“…In the gastrointestinal system, it comprises a range of diseases, from the less defined irritable bowel syndrome to full impairment of intestinal function in inflammatory bowel disease (IBD) [60]. A leaky barrier is gradually being appointed as part of these inflammatory and infectious diseases [42]. Apostolou et al [42] used the proprietary Human Emulation System Organ-on-a-chip by Emulate® to develop their system: Colon Intestine-Chip.…”