A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal Permeability
| Autor: | Rohit A. Panchakshari, Athanasia Apostolou, Cosmas Giallourakis, Galeb Abu-Ali, Antara Banerjee, Tengku Ibrahim Maulana, Raymond Luc, Gauri Kulkarni, Carolina Lucchesi, Jordan Kerns, Katia Karalis, Alexandra Dimitriou, Maria D. Paraskevopoulou, Magdalena Kasendra, Elias S. Manolakos, Lorna Ewart, Geraldine A. Hamilton, Bertram Bleck, Dimitris V. Manatakis |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
EdU
5-ethynyl-2’-deoxyuridine Cell Culture Techniques RC799-869 Wnt wingless-related integration site IEC intestinal epithelial cell Interleukin 22 ZO-1 zonula occludens-1 IL interleukin 6 Organ-on-Chip Leaky Gut Lab-On-A-Chip Devices Intestinal Mucosa Receptor Original Research DKK1 Dickkopf-related protein 1 IBD inflammatory bowel disease cHIMEC colonic human intestinal microvascular endothelial cell Gastroenterology Diseases of the digestive system. Gastroenterology Intestinal epithelium NHS N-hydroxysuccinimide Cell biology ECM extracellular matrix Organoids STAT signal transducer and activator of transcription TNFα tumor necrosis factor α medicine.anatomical_structure TJ tight junction Cellular Microenvironment PDMS polydimethylsiloxane qPCR quantitative polymerase chain reaction Endothelium Colon PBS phosphate-buffered saline Biology Permeability In vivo medicine Organoid Humans DGE differential gene expression IFNγ interferon-γ GO gene ontology Intestinal permeability Hepatology Mechanism (biology) Gene Expression Profiling Interleukins RNA-seq RNA sequencing Computational Biology medicine.disease IL22BP interleukin 22 binding protein Papp apparent permeability Gene Expression Regulation Transcriptome Biomarkers |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology Cellular and Molecular Gastroenterology and Hepatology, Vol 12, Iss 5, Pp 1719-1741 (2021) |
| ISSN: | 2352-345X |
| Popis: | 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 (Emulate, Inc, Boston, MA) 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, 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 cytokine-induced barrier disruption. Results Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice. Conclusions We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers. Graphical abstract |
| Databáze: | OpenAIRE |
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