Periodic Reporting for period 3 - GutHormones (Understanding the Biology of Human Enteroendocrine Cells)
Okres sprawozdawczy: 2023-10-01 do 2025-03-31
The intestinal tract is a complex organ system in which a single layer of specialized epithelium, replaced every week by the stem cells, performs its primary functions of digestion, absorption, protection and excretion. The intestinal epithelium consists, therefore, of different epithelial cells that secrete digestive enzymes and mucus (goblet cells), anti-bacterial agents (Paneth cells), absorb food particles (enterocytes) or produce hormones (entero-endocrine cells).
The rare EECs, also present in the stomach and colon, are subclassified by the (>20) different hormones they produce. Generally, EECs sense intestinal content and regulate gastrointestinal activity, systemic metabolism and food intake. While one EEC hormone, GLP1, has been developed into a highly successful diabetes drug, a paucity of laboratory models has complicated human EEC studies.
We have previously identified the adult intestinal stem cell and established conditions to grow ‘gut organoids’ (mini-organs in a dish) from these stem cells. These organoids have been used to resolve the complex EEC subtype hierarchy. Moreover, we developed protocols in which gut organoids can be manipulated to contain all EEC lineages and can be genetically modified using CRISPR technology. E.g. individual hormones can be tagged by a fluorescent marker. These cells can subsequently isolated and/or analyzed on basis of the fluorescent marker.
The ERC grant GutHormones exploits the technologies described above to generate a detailed picture of the biology of EECs, their triggers, their interconnectivity and their products.
Successful completion of this project will generate a comprehensive characterization of EECs. This will inspire new therapeutic approaches to reshape the EEC landscape for the treatment of metabolic diseases (obesity, diabetes, fatty liver disease, atherosclerosis).
The rare EECs, also present in the stomach and colon, are subclassified by the (>20) different hormones they produce. Generally, EECs sense intestinal content and regulate gastrointestinal activity, systemic metabolism and food intake. While one EEC hormone, GLP1, has been developed into a highly successful diabetes drug, a paucity of laboratory models has complicated human EEC studies.
We have previously identified the adult intestinal stem cell and established conditions to grow ‘gut organoids’ (mini-organs in a dish) from these stem cells. These organoids have been used to resolve the complex EEC subtype hierarchy. Moreover, we developed protocols in which gut organoids can be manipulated to contain all EEC lineages and can be genetically modified using CRISPR technology. E.g. individual hormones can be tagged by a fluorescent marker. These cells can subsequently isolated and/or analyzed on basis of the fluorescent marker.
The ERC grant GutHormones exploits the technologies described above to generate a detailed picture of the biology of EECs, their triggers, their interconnectivity and their products.
Successful completion of this project will generate a comprehensive characterization of EECs. This will inspire new therapeutic approaches to reshape the EEC landscape for the treatment of metabolic diseases (obesity, diabetes, fatty liver disease, atherosclerosis).
With financial support from the EU (ERC grant GutHormones), we will generate a detailed picture of the biology of the hormone-producing entero-endocrine cells (EECs), their triggers, their interconnectivity and their products. This ultimate aim is to provide a deep understanding of EECs, potentially leading to new treatments for obesity, diabetes, and other metabolic diseases by targeting these hormone-producing cells.
To reach our aim, we have built a platform where all hormone-producing human EECs of the small intestine, stomach and colon can be induced in high numbers in organoids (mini organs in a dish). Additionally, EECs could be isolated and analysed based on fluorescent tags, which we genetically introduced using the novel CRISPR technology. With this, we successfully completed the gastrointestinal human EEC atlases and hormone-tagged organoid collection.
Using these organoids in combination with the deletion of many different receptors, we have identified the signals which are critical regulators of setting the fate of the different EECs.
We performed experiments to resolve which luminal receptors and ligands allow EECs to sample and evaluate the content of the intestinal lumen. The obtained results, describing and functional validating the EEC ‘sensorome’, will act as a starting point for the rational design of therapeutics that target these epithelial cells specifically.
EECs express numerous receptors for hormones secreted by the same endocrine cells. To start assessing the interplay between EECs and other epithelial and non-epithelial lineages, we are constructing a set of organoids in which synthetic receptors allow activation of individual EEC subsets expressing the pertinent hormone, causing all hormone products to be secreted. These innovative models will be used to study the effect of secretion of EEC subsets on other EEC subtypes, as well as on other lineages and distant organs (hepatocytes, islets).
To reach our aim, we have built a platform where all hormone-producing human EECs of the small intestine, stomach and colon can be induced in high numbers in organoids (mini organs in a dish). Additionally, EECs could be isolated and analysed based on fluorescent tags, which we genetically introduced using the novel CRISPR technology. With this, we successfully completed the gastrointestinal human EEC atlases and hormone-tagged organoid collection.
Using these organoids in combination with the deletion of many different receptors, we have identified the signals which are critical regulators of setting the fate of the different EECs.
We performed experiments to resolve which luminal receptors and ligands allow EECs to sample and evaluate the content of the intestinal lumen. The obtained results, describing and functional validating the EEC ‘sensorome’, will act as a starting point for the rational design of therapeutics that target these epithelial cells specifically.
EECs express numerous receptors for hormones secreted by the same endocrine cells. To start assessing the interplay between EECs and other epithelial and non-epithelial lineages, we are constructing a set of organoids in which synthetic receptors allow activation of individual EEC subsets expressing the pertinent hormone, causing all hormone products to be secreted. These innovative models will be used to study the effect of secretion of EEC subsets on other EEC subtypes, as well as on other lineages and distant organs (hepatocytes, islets).