A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal Permeability

Abstract

Colon Intestine-Chip Q9 (Emulate, Inc, Boston, MA), seeded with human colon crypt-derived epithelial and primary microvascular endothelial cells, was built to investigate leaky gut in human beings. This ex vivo platform recapitulated the effects of proinflammatory cytokines in the intestinal epithelial barrier and identified novel mechanisms of action. BACKGROUND & AIMS:The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip Q10 platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity. METHODS:We combined the human biopsy-derived leucine-rich repeat-containing G-protein-coupled receptor 5-positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate Q11 , Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokineinduced barrier disruption. RESULTS:Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment

Organoids as tools for fundamental discovery and translation—a Keystone Symposia report

“…The addition of IL‐22, which has been implicated in the pathogenesis of IBD and is generally thought to support the epithelial barrier, 17–19 showed, as anticipated, that IL‐22 induced STAT3 activation in a concentration‐ and time‐dependent manner. However, unexpectedly, IL‐22 compromised epithelial morphology and induced apoptosis 15 . These data show the importance of using more in vivo‐ like and species‐specific experimental models to understand human diseases.…”

“…She showed that coculture of endothelial and epithelial cells recreated human colon physiology more closely than do classical organoids, with enhanced formation of a tight junction network and epithelial polarity (Figure 1). Applying mechanical force to the Colon‐Chips altered transcription, increasing the expression of genes involved in ion, lipoprotein, and water transportation 15 . Apostolou also showed how the Colon Intestine‐Chip responds to factors known to play a role in IBD while providing additional insights into their effects on the colon.…”

“…However, unexpectedly, IL-22 compromised epithelial morphology and induced apoptosis. 15 These data show the importance of using more in vivo-like and species-specific experimental models to understand human diseases.…”

“…Applying mechanical force to the Colon-Chips altered transcription, increasing the expression of genes involved in ion, lipoprotein, and water transportation. 15 Apostolou also showed how the Colon Intestine-Chip responds to factors known to play a role in IBD while providing additional insights into their effects on the colon. The addition of IFN-γ, a cytokine that disrupts the epithelial barrier, 16 resulted in a time-dependent collapse of the epithelial barrier, compromised epithelial morphology, disruption of tight junctions, and restructuring of the actin cytoskeleton.…”

Gut-on-a-chip models for dissecting the gut microbiology and physiology

“… 71,92 At the transcriptome level, peristalsis enhanced gene expressions related to ion, lipoprotein, and water transportation, as evidenced in the analysis of organoid-derived epithelial cells on chip. 110 …”