Journal
CELLULAR AND MOLECULAR GASTROENTEROLOGY AND HEPATOLOGY
Volume 5, Issue 3, Pages 333-352Publisher
ELSEVIER INC
DOI: 10.1016/j.jcmgh.2017.12.013
Keywords
Three-Dimensional; Keratinocytes; Eosinophilic Esophagitis; Squamous Cell Differentiation
Categories
Funding
- NATIONAL CANCER INSTITUTE [F31CA174176, U54CA163004, P01CA098101] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [U54AI117804] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [K01DK103953, F32DK100088, P30DK050306, K01DK100485, R01DK114436, R03DK114463, K08DK106444] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES [P30ES013508] Funding Source: NIH RePORTER
- NCI NIH HHS [U54 CA163004, F31 CA174176, P01 CA098101] Funding Source: Medline
- NIAID NIH HHS [U54 AI117804] Funding Source: Medline
- NIDDK NIH HHS [R03 DK114463, K01 DK103953, F32 DK100088, K01 DK100485, K08 DK106444, R01 DK114436, P30 DK050306] Funding Source: Medline
- NIEHS NIH HHS [P30 ES013508] Funding Source: Medline
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BACKGROUND & AIMS: Aberrations in the esophageal proliferation-differentiation gradient are histologic hallmarks in eosinophilic esophagitis (EoE) and gastroesophageal reflux disease. A reliable protocol to grow 3-dimensional (3D) esophageal organoids is needed to study esophageal epithelial homeostasis under physiological and pathologic conditions. METHODS: We modified keratinocyte-serum free medium to grow 3D organoids from endoscopic esophageal biopsies, immortalized human esophageal epithelial cells, and murine esophagi. Morphologic and functional characterization of 3D organoids was performed following genetic and pharmacologic modifications or exposure to EoE-relevant cytokines. The Notch pathway was evaluated by transfection assays and by gene expression analyses in vitro and in biopsies. RESULTS: Both murine and human esophageal 3D organoids displayed an explicit proliferation-differentiation gradient. Notch inhibition accumulated undifferentiated basal keratinocytes with deregulated squamous cell differentiation in organoids. EoE patient-derived 3D organoids displayed normal epithelial structure ex vivo in the absence of the EoE inflammatory milieu. Stimulation of esophageal 3D organoids with EoE-relevant cytokines resulted in a phenocopy of Notch inhibition in organoid 3D structures with recapitulation of reactive epithelial changes in EoE biopsies, where Notch3 expression was significantly decreased in EoE compared with control subjects. CONCLUSIONS: Esophageal 3D organoids serve as a novel platform to investigate regulatory mechanisms in squamous epithelial homeostasis in the context of EoE and other diseases. Notch-mediated squamous cell differentiation is suppressed by cytokines known to be involved in EoE, suggesting that this may contribute to epithelial phenotypes associated with disease. Genetic and pharmacologic manipulations establish proof of concept for the utility of organoids for future studies and personalized medicine in EoE and other esophageal diseases.
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