Modeling epithelial homeostasis and perturbation in three-dimensional human esophageal organoids

Shimonosono, M.; Morimoto, M.; Hirose, W.; Tomita, Y.; Matsuura, N.; Flashner, S.; Ebadi, M. S.; Okayasu, E. H.; Lee, C. Y.; Britton, W. R.; Martin, C.; Wuertz, B. R.; Parikh, A. S.; Sachdeva, U. M.; Ondrey, F. G.; Atigadda, V. R.; Elmets, C. A.; Abrams, J.; Muir, A. B.; Klein-Szanto, A. J.; Weinberg, K. I.; Momen-Heravi, F.; Nakagawa, H.

doi:10.1101/2024.05.20.595023

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bioRxiv · 10.1101/2024.05.20.595023

Modeling epithelial homeostasis and perturbation in three-dimensional human esophageal organoids

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Abstract

BackgroundEsophageal organoids from a variety of pathologies including cancer are grown in Advanced Dulbeccos Modified Eagle Medium-Nutrient Mixture F12 (hereafter ADF). However, the currently available ADF-based formulations are suboptimal for normal human esophageal organoids, limiting the ability to compare normal esophageal organoids with those representing a given disease state. MethodsWe have utilized immortalized normal human esophageal epithelial cell (keratinocyte) lines EPC1 and EPC2 and endoscopic normal esophageal biopsies to generate three-dimensional (3D) organoids. To optimize ADF-based medium, we evaluated the requirement of exogenous epidermal growth factor (EGF) and inhibition of transforming growth factor-(TGF)-{beta} receptor-mediated signaling, both key regulators of proliferation of human esophageal keratinocytes. We have modeled human esophageal epithelial pathology by stimulating esophageal 3D organoids with interleukin (IL)-13, an inflammatory cytokine, or UAB30, a novel pharmacological activator of retinoic acid signaling. ResultsThe formation of normal human esophageal 3D organoids was limited by excessive EGF and intrinsic TGF{beta} receptor-mediated signaling. In optimized HOME0, normal human esophageal organoid formation was improved, whereas IL-13 and UAB30 induced epithelial changes reminiscent of basal cell hyperplasia, a common histopathologic feature in broad esophageal disease conditions including eosinophilic esophagitis. Conclusions: HOME0 allows modeling of the homeostatic differentiation gradient and perturbation of the human esophageal epithelium while permitting a comparison of organoids from mice and other organs grown in ADF-based media.

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Shimonosono, M., Morimoto, M., Hirose, W., Tomita, Y., Matsuura, N., Flashner, S., Ebadi, M. S., Okayasu, E. H., Lee, C. Y., Britton, W. R., Martin, C., Wuertz, B. R., Parikh, A. S., Sachdeva, U. M., Ondrey, F. G., Atigadda, V. R., Elmets, C. A., Abrams, J., Muir, A. B., Klein-Szanto, A. J., Weinberg, K. I., Momen-Heravi, F., Nakagawa, H.. 2024-05-21. Modeling epithelial homeostasis and perturbation in three-dimensional human esophageal organoids. https://doi.org/10.1101/2024.05.20.595023

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