ArticleCellular and molecular gastroenterology and hepatology2019
Human Intestinal Enteroids With Inducible Neurogenin-3 Expression as a Novel Model of Gut Hormone Secretion.
Article in Cellular and molecular gastroenterology and hepatology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 56 papers.
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56 citing papers in PubMed, 82 citations in OpenAlex.
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- Microbial stimulation of oxytocin release from the intestinal epithelium via secretin signaling.Gut microbes · 2023Article
- Human intestinal organoids from Cronkhite-Canada syndrome patients reveal link between serotonin and proliferation.The Journal of clinical investigation · 2023Article
- Human intestinal organoids as models to study enteric bacteria and viruses.Current opinion in microbiology · 2023Review
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- Microbial stimulation of oxytocin release from the intestinal epithelium via secretin signaling.bioRxiv : the preprint server for biology · 2023Article
- The possible role of gut microbiota dysbiosis in the pathophysiology of delirium in older persons.Microbiome research reports · 2023Review
- Co-culturing Human Intestinal Enteroid Monolayers with Innate Immune Cells.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Gastrointestinal organoids in the study of viral infections.American journal of physiology. Gastrointestinal and liver physiology · 2023Review
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9 authors at 1 institution in 1 country.
Funding
Abstract
BACKGROUND &
aimsEnteroendocrine cells (EECs) are specialized epithelial cells that produce molecules vital for intestinal homeostasis, but because of their limited numbers, in-depth functional studies have remained challenging. Human intestinal enteroids (HIEs) that are derived from intestinal crypt stem cells are biologically relevant in an in vitro model of the intestinal epithelium. HIEs contain all intestinal epithelial cell types; however, similar to the intestine, HIEs spontaneously produce few EECs, which limits their study.
methodsTo increase the number of EECs in HIEs, we used lentivirus transduction to stably engineer jejunal HIEs with doxycycline-inducible expression of neurogenin-3 (NGN3), a transcription factor that drives EEC differentiation (tetNGN3-HIEs). We examined the impact of NGN3 induction on EECs by quantifying the increase in the enterochromaffin cells and other EEC subtypes. We functionally assessed secretion of serotonin and EEC hormones in response to norepinephrine and rotavirus infection.
resultsTreating tetNGN3-HIEs with doxycycline induced a dose-dependent increase of chromogranin A (ChgA)-positive and serotonin-positive cells, showing increased enterochromaffin cell differentiation. Despite increased ChgA-positive cells, other differentiated cell types of the epithelium remained largely unchanged by gene expression and immunostaining. RNA sequencing of doxycycline-induced tetNGN3-HIEs identified increased expression of key hormones and enzymes associated with several other EEC subtypes. Doxycycline-induced tetNGN3-HIEs secreted serotonin, monocyte chemoattractant protein-1, glucose-dependent insulinotropic peptide, peptide YY, and ghrelin in response to norepinephrine and rotavirus infection, further supporting the presence of multiple EEC types.
conclusionsWe have combined HIEs and inducible-NGN3 expression to establish a flexible in vitro model system for functional studies of EECs in enteroids and advance the molecular and physiological investigation of EECs.
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