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2022
Villegas-Novoa, Cecilia; Wang, Yuli; Sims, Christopher E.; Allbritton, Nancy L.
Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion Journal Article
In: Development, vol. 94, no. 27, pp. 9648–9655, 2022, ISSN: 1520-6882.
Abstract | Links | BibTeX | Tags: Diabetes Mellitus, Enteroendocrine Cells, Glucagon-Like Peptide 1, Intestinal Mucosa, L Cells
@article{villegas-novoa_development_2022,
title = {Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion},
author = {Cecilia Villegas-Novoa and Yuli Wang and Christopher E. Sims and Nancy L. Allbritton},
doi = {10.1021/acs.analchem.2c00912},
issn = {1520-6882},
year = {2022},
date = {2022-07-12},
urldate = {2022-07-12},
journal = {Development},
volume = {94},
number = {27},
pages = {9648–9655},
abstract = {Type 2 diabetes mellitus is a chronic disease associated with obesity and dysregulated human feeding behavior. The hormone glucagon-like peptide 1 (GLP-1), a critical regulator of body weight, food intake, and blood glucose levels, is secreted by enteroendocrine L-cells. The paucity of L-cells in primary intestinal cell cultures including organoids and monolayers has made assays of GLP-1 secretion from primary human cells challenging. In the current paper, an analytical assay pipeline consisting of an optimized human intestinal tissue construct enriched in L-cells paired with standard antibody-based GLP-1 assays was developed to screen compounds for the development of pharmaceuticals to modulate L-cell signaling. The addition of the serotonin receptor agonist Bimu 8, optimization of R-spondin and Noggin concentrations, and utilization of vasoactive intestinal peptide (VIP) increased the density of L-cells in a primary human colonic epithelial monolayer. Additionally, the incorporation of an air-liquid interface culture format increased the L-cell number so that the signal-to-noise ratio of conventional enzyme-linked immunoassays could be used to monitor GLP-1 secretion in compound screens. To demonstrate the utility of the optimized analytical method, 21 types of beverage sweeteners were screened for their ability to stimulate GLP-1 secretion. Stevioside and cyclamate were found to be the most potent inducers of GLP-1 secretion. This platform enables the quantification of GLP-1 secretion from human primary L-cells and will have broad application in understanding L-cell formation and physiology and will improve the identification of modulators of human feeding behavior.},
keywords = {Diabetes Mellitus, Enteroendocrine Cells, Glucagon-Like Peptide 1, Intestinal Mucosa, L Cells},
pubstate = {published},
tppubtype = {article}
}
Type 2 diabetes mellitus is a chronic disease associated with obesity and dysregulated human feeding behavior. The hormone glucagon-like peptide 1 (GLP-1), a critical regulator of body weight, food intake, and blood glucose levels, is secreted by enteroendocrine L-cells. The paucity of L-cells in primary intestinal cell cultures including organoids and monolayers has made assays of GLP-1 secretion from primary human cells challenging. In the current paper, an analytical assay pipeline consisting of an optimized human intestinal tissue construct enriched in L-cells paired with standard antibody-based GLP-1 assays was developed to screen compounds for the development of pharmaceuticals to modulate L-cell signaling. The addition of the serotonin receptor agonist Bimu 8, optimization of R-spondin and Noggin concentrations, and utilization of vasoactive intestinal peptide (VIP) increased the density of L-cells in a primary human colonic epithelial monolayer. Additionally, the incorporation of an air-liquid interface culture format increased the L-cell number so that the signal-to-noise ratio of conventional enzyme-linked immunoassays could be used to monitor GLP-1 secretion in compound screens. To demonstrate the utility of the optimized analytical method, 21 types of beverage sweeteners were screened for their ability to stimulate GLP-1 secretion. Stevioside and cyclamate were found to be the most potent inducers of GLP-1 secretion. This platform enables the quantification of GLP-1 secretion from human primary L-cells and will have broad application in understanding L-cell formation and physiology and will improve the identification of modulators of human feeding behavior.