Modeling Epithelial Homeostasis and Reactive Epithelial Changes in Human and Murine Three‐Dimensional Esophageal Organoids
Year: 2020
Type: article
Abstract: The homeostatic proliferation-differentiation gradient in the esophageal epithelium is perturbed under inflammatory disease conditions such as gastroesophageal reflux disease and eosinophilic esophagitis. Herein we describe the protocols for rapid generation (<14 days) and characterization of single-cell-derived, three-dimensional (3D) esophageal organoids from human subjects and mice with normal esophageal mucosa or inflammatory disease conditions. While 3D organoids recapitulate normal epithelial renewal, proliferation, and differentiation, non-cell autonomous reactive epithelial changes under inflammatory conditions are evaluated in the absence of the inflammatory milieu. Reactive epithelial changes are reconstituted upon exposure to exogenous recombinant cytokines. These changes are modulated pharmacologically or genetically ex vivo. Molecular, structural, and functional changes are characterized by morphology, flow cytometry, biochemistry, and gene expression analyses. Esophageal 3D organoids can be translated for the development of personalized medicine in assessment of individual cytokine sensitivity and molecularly targeted therapeutics in esophagitis patients © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Generation of esophageal organoids from biopsy or murine esophageal epithelial sheets Basic Protocol 2: Propagation and cryopreservation of esophageal organoids Basic Protocol 3: Harvesting of esophageal organoids for RNA isolation, immunohistochemistry, and evaluation of 3D architecture Basic Protocol 4: Modeling of reactive epithelium in esophageal organoids.
DOI: 10.1002/cpsc.106
Authors: Hiroshi Nakagawa, Yuta Kasagi, Tatiana A. Karakasheva, Takeo Hara, Bailey Aaron, Masataka Shimonosono, Takashi Kijima, Veronique Giroux, Dominique Bailey, Benjamin Wilkins, Julian A. Abrams, Gary W. Falk, Seema S. Aceves, Jonathan M. Spergel, Kathryn E. Hamilton, Kelly A. Whelan, Amanda B. Muir
Language: English
Topic: Eosinophilic Esophagitis
Subfield: Gastroenterology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Funders: National Science Foundation, Children's Hospital of Philadelphia, American Partnership for Eosinophilic Disorders, North American Society for Pediatric Gastroenterology, Hepatology and Nutrition, National Institutes of Health, National Institute on Alcohol Abuse and Alcoholism, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Environmental Health Sciences, National Center for Advancing Translational Sciences
Awards: Contribution of rigid matrix to allergic eosinophilic esophagitis pathogenesis, The Role of the Microenvironment in Barrett's Esophagus, Consortium of Eosinophilic Gastrointestinal Disease Researchers, Mechanisms of Esophageal Carcinogenesis, The functional role of mitophagy in esophageal cell biology, Lysyl oxidase mediated fibrosis in eosinophilic esophagitis, G2312078907, G2626970174, G2969096386, Mentoring Patient Oriented Research in Allergic Eosinophilic Gastrointestinal Disorders, Autophagy regulation of esophageal basal cell dynamics, G3148304603, Autophagy and esophageal tissue remodeling in EoE, G397555987, G4021028342, G4041853508, Esophageal tissue aging under homeostatic and inflammatory conditions, The Role of Lysyl Oxidase in Epithelial Differentiation in Eosinophilic Esophagitis, G7668806916, The role of IMP1 mRNA binding protein in intestinal epithelial biology, G776886754, Mechanism of Respiratory System-Esophageal Separation, G8963766838, Center of Excellence in Environmental Toxicology, Data Management and Coordinating Center: Rare Diseases Clinical Research Network, Aldh2 and epithelial homeostasis and pathobiology, Multidisciplinary Training in Translational Gastrointestinal and Liver Research, Center for Molecular Studies in Digestive and Liver Diseases, Cancer Center Support Grant, IMP1 REGULATION OF AUTOPHAGY IN THE INTESTINAL EPITHELIUM
OpenAlex ID: W3007918331