{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T12:32:22Z","timestamp":1791635542078,"version":"4.3.4"},"reference-count":47,"publisher":"Wiley","issue":"24","license":[{"start":{"date-parts":[[2024,12,5]],"date-time":"2024-12-05T00:00:00Z","timestamp":1733356800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2025,12]]},"abstract":"\n Abstract<\/jats:title>\n \n Gut hormones control intestinal function, metabolism and appetite, and have been harnessed therapeutically to treat type 2 diabetes and obesity. Our understanding of the enteroendocrine axis arises largely from animal studies, but intestinal organoid models make it possible to identify, genetically modify and purify human enteroendocrine cells (EECs). This study aimed to map human EECs using single\u2010cell RNA sequencing. Organoids derived from human duodenum and ileum were genetically modified using CRISPR\u2010Cas9 to express the fluorescent protein Venus driven by the chromogranin\u2010A promoter. Fluorescent cells from CHGA\u2010Venus organoids were purified by flow cytometry and analysed by 10X single\u2010cell RNA sequencing. Cluster analysis separated EEC populations, allowing an examination of differentially expressed hormones, nutrient\u2010sensing machinery, transcription factors and exocytotic machinery. Bile acid receptor\n GPBAR1<\/jats:italic>\n was most highly expressed in L\u2010cells (producing glucagon\u2010like peptide 1 and peptide YY), long\u2010chain fatty acid receptor\n FFAR1<\/jats:italic>\n was highest in I\u2010cells (cholecystokinin), K\u2010cells (glucose\u2010dependent insulinotropic polypeptide) and L\u2010cells, short\u2010chain fatty acid receptor\n FFAR2<\/jats:italic>\n was highest in ileal L\u2010cells and enterochromaffin cells, olfactory receptor\n OR51E1<\/jats:italic>\n was notably expressed in ileal enterochromaffin cells, and the glucose\u2010sensing sodium glucose cotransporter\n SLC5A1<\/jats:italic>\n was highly and differentially expressed in K\u2010 and L\u2010cells, reflecting their known responsiveness to ingested glucose. The organoid EEC atlas was merged with published data from human intestine and organoids, with good overlap between enteroendocrine datasets. Understanding the similarities and differences between human EEC types will facilitate the development of drugs targeting the enteroendocrine axis for the treatment of conditions such as diabetes, obesity and intestinal disorders.\n \n \n image<\/jats:alt-text>\n <\/jats:graphic>\n <\/jats:boxed-text>\n <\/jats:p>\n <\/jats:sec>\n \n Key points<\/jats:title>\n \n \n \n Gut hormones regulate intestinal function, nutrient homeostasis and metabolism and form the basis of the new classes of drugs for obesity and diabetes.<\/jats:p>\n <\/jats:list-item>\n \n As enteroendocrine cells (EECs) comprise only \u223c1% of the intestinal epithelium, they are under\u2010represented in current single\u2010cell atlases.<\/jats:p>\n <\/jats:list-item>\n \n To identify, compare and characterise human EECs we generated chromogranin\u2010A labelled organoids from duodenal and ileal biopsies by CRISPR\u2010Cas9.<\/jats:p>\n <\/jats:list-item>\n \n Fluorescent chromogranin\u2010A positive EECs were purified and analysed by single\u2010cell RNA sequencing, revealing predominant cell clusters producing different gut hormones.<\/jats:p>\n <\/jats:list-item>\n \n Cell clusters exhibited differential expression of nutrient\u2010sensing machinery including bile acid receptors, long\u2010 and short\u2010chain fatty acid receptors and glucose transporters.<\/jats:p>\n <\/jats:list-item>\n \n Organoid\u2010derived EECs mapped well onto data from native intestinal cell populations, extending coverage of EECs.<\/jats:p>\n <\/jats:list-item>\n <\/jats:list>\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1113\/jp287463","type":"journal-article","created":{"date-parts":[[2024,12,6]],"date-time":"2024-12-06T02:28:54Z","timestamp":1733452134000},"page":"7751-7763","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Single\u2010cell transcriptomics of human organoid\u2010derived enteroendocrine cell populations from the small intestine"],"prefix":"10.1113","volume":"603","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5035-6828","authenticated-orcid":false,"given":"Christopher A.","family":"Smith","sequence":"first","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4880-6455","authenticated-orcid":false,"given":"Van B.","family":"Lu","sequence":"additional","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5643-0195","authenticated-orcid":false,"given":"Rula","family":"Bany Bakar","sequence":"additional","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9201-0860","authenticated-orcid":false,"given":"Emily","family":"Miedzybrodzka","sequence":"additional","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam","family":"Davison","sequence":"additional","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah","family":"Goldspink","sequence":"additional","affiliation":[{"name":"Institute of Metabolic Science and MRC Metabolic Diseases Unit University of Cambridge, Addenbrooke's Hospital Cambridge 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