Exploring the function and sensing mechanisms of human enteroendocrine cells using intestinal organoids
type: CreativeWork
description: The gut is the largest endocrine organ in our body thanks to the presence of hormone-releasing enteroendocrine cells (EECs), which represent only around 1% of the total gut epithelium. Gut hormones are released in response to cues of different natures to regulate several physiological events, such as nutrient digestion and absorption, intestinal motility, food intake, inflammation, and energy homeostasis. This thesis aimed to characterise hormone-releasing sensing mechanisms adopted by two important types of EECs: K-cells, which secrete glucose-dependent insulinotropic polypeptide (GIP), and enterochromaffin (EC) cells, which secrete serotonin, or 5-hydroxytryptamine. GIP, largely known as one of the two gut-derived insulinotropic incretin hormones, alongside with glucagon-like peptide 1 (GLP-1), has recently regained popularity in the field of metabolic research due to its newfound relevance as an anorexigenic hormone in rodent models and the advent of weight loss-inducing GLP1R/GIPR dual incretin mimetics. 5-HT is a monoamine neurotransmitter and non-canonical gut hormone that has a wide impact on the gut and whole-body physiology. Most studies exploring EEC function have used either animal models or 2D cell lines. However, the development of human gut organoids, 3D structures that recapitulate the intestinal epithelial complexities, and gene-editing tools, such as CRISPR-Cas9, paired with the establishment of robust characterisation assays, have allowed us, for the first time, to explore human EEC function and hormone secretory mechanism in depth. Using CRISPR-Cas9 homology directed repair (HDR), we fluorescently labelled K-cells and EC cells in patient-derived duodenal organoids. These transgenic reporter lines were employed to characterise human EEC responsiveness and function by performing fluorescence-activated cell sorting (FACS), bulk RNA sequencing, hormone secretion assays and live single-cell Ca2+ and cAMP imaging. cAMP imaging was possible through the g…
abstract: The gut is the largest endocrine organ in our body thanks to the presence of hormone-releasing enteroendocrine cells (EECs), which represent only around 1% of the total gut epithelium. Gut hormones are released in response to cues of different natures to regulate several physiological events, such as nutrient digestion and absorption, intestinal motility, food intake, inflammation, and energy homeostasis. This thesis aimed to characterise hormone-releasing sensing mechanisms adopted by two important types of EECs: K-cells, which secrete glucose-dependent insulinotropic polypeptide (GIP), and enterochromaffin (EC) cells, which secrete serotonin, or 5-hydroxytryptamine. GIP, largely known as one of the two gut-derived insulinotropic incretin hormones, alongside with glucagon-like peptide 1 (GLP-1), has recently regained popularity in the field of metabolic research due to its newfound relevance as an anorexigenic hormone in rodent models and the advent of weight loss-inducing GLP1R/GIPR dual incretin mimetics. 5-HT is a monoamine neurotransmitter and non-canonical gut hormone that has a wide impact on the gut and whole-body physiology. Most studies exploring EEC function have used either animal models or 2D cell lines. However, the development of human gut organoids, 3D structures that recapitulate the intestinal epithelial complexities, and gene-editing tools, such as CRISPR-Cas9, paired with the establishment of robust characterisation assays, have allowed us, for the first time, to explore human EEC function and hormone secretory mechanism in depth. Using CRISPR-Cas9 homology directed repair (HDR), we fluorescently labelled K-cells and EC cells in patient-derived duodenal organoids. These transgenic reporter lines were employed to characterise human EEC responsiveness and function by performing fluorescence-activated cell sorting (FACS), bulk RNA sequencing, hormone secretion assays and live single-cell Ca2+ and cAMP imaging. cAMP imaging was possible through the g…
author: Guccio, Nunzio
language: eng
keywords: Enteroendocrine, Hormone, Intestinal, Metabolic disease, Organoids
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/add147ae-ee23-41e1-ae94-af4a8129c855/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2024-04-18
date modified: 2026-07-22T22:24:14Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.111638