Enteroendocrine cells (EECs) sense intestinal content and release hormones to regulate gastrointestinal activity, systemic metabolism, and food intake. Little is known about the molecular make-up of human EEC subtypes and the regulated secretion of individual hormones. Here, we describe an organoid-based platform for functional studies of human EECs. EEC formation is induced in vitro by transient expression of NEUROG3. A set of gut organoids was engineered in which the major hormones are fluorescently tagged. A single-cell mRNA atlas was generated for the different EEC subtypes, and their secreted products were recorded by mass-spectrometry. We note key differences to murine EECs, including hormones, sensory receptors, and transcription factors. Notably, several hormone-like molecules were identified. Inter-EEC communication is exemplified by secretin-induced GLP-1 secretion. Indeed, individual EEC subtypes carry receptors for various EEC hormones. This study provides a rich resource to study human EEC development and function.
Keywords: CRISPR-mediated knockin; enteroendocrine cell; fluorescent tagging; hormone; human; intestine; organoids; proteomics; secretomics; single-cell atlas.
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Conflict of interest statement
Declaration of Interests H.C. is inventor on several patents related to organoid technology; his full disclosure is given at https://www.uu.nl/staff/JCClevers/. H.C. is founder of OrganoidZ, which employs organoids for drug development. J.B., J.P., and H.C. are inventors on patents related to this work.
Deriving functional human enteroendocrine cells from pluripotent stem cells.Development. 2018 Oct 1;145(19):dev165795. doi: 10.1242/dev.165795. Development. 2018. PMID: 30143540 Free PMC article.
Comparison of enteroendocrine cells and pancreatic β-cells using gene expression profiling and insulin gene methylation.PLoS One. 2018 Oct 31;13(10):e0206401. doi: 10.1371/journal.pone.0206401. eCollection 2018. PLoS One. 2018. PMID: 30379923 Free PMC article.
Human Intestinal Enteroids With Inducible Neurogenin-3 Expression as a Novel Model of Gut Hormone Secretion.Cell Mol Gastroenterol Hepatol. 2019;8(2):209-229. doi: 10.1016/j.jcmgh.2019.04.010. Epub 2019 Apr 25. Cell Mol Gastroenterol Hepatol. 2019. PMID: 31029854 Free PMC article.
Enteroendocrine cells: a review of their role in brain-gut communication.Neurogastroenterol Motil. 2016 May;28(5):620-30. doi: 10.1111/nmo.12754. Epub 2015 Dec 21. Neurogastroenterol Motil. 2016. PMID: 26691223 Free PMC article. Review.
Evolving Concepts and Translational Relevance of Enteroendocrine Cell Biology.J Clin Endocrinol Metab. 2016 Mar;101(3):778-86. doi: 10.1210/jc.2015-3449. Epub 2015 Dec 9. J Clin Endocrinol Metab. 2016. PMID: 26649620 Review.