Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion

Abstract

Type 2 diabetes mellitus is a chronic disease associated with obesity and dysregulated human feeding behavior. The hormone glucagonlike 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 Lcells 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.

Mechano-regulation of GLP-1 production by Piezo1 in intestinal L cells

“…Whether Piezo1 plays the same role in human L cells awaits to be investigated. A number of studies have generated L cells culture from human intestinal organoid culture or human intestinal stem cell monolayer culture by manipulating the growth factors in the media (Goldspink et al, 2020; Petersen et al, 2014; Villegas-Novoa et al, 2022). It is worthy to validate our finding in human L cells in order to prove its translational potential in T2DM treatment.…”

Separation of the effects of small intestinal microbiome-diet interactions on human gut hormone secretion

“…Lastly, we used a model of gut hormone secretion from an engineered jejunal organoid line (Chang-Graham et al, 2019). If hormone secretion from a non-engineered organoid line is needed, changes to the organoid growth media could be employed to enhance the abundance of enteroendocrine cells (Fujii et al, 2018; Villegas-Novoa et al, 2022; Zeve et al, 2022).…”

Applications of human organoids in the personalized treatment for digestive diseases

“… 391 The serotonin receptor agonist Bimu8 could increase the density of L-cells in primary human colonic organoids. 392 Pretreatment with L-arginine induces the stemness of the stem cell pool and preserves the gut response to TNF-α and 5-fluorouracil. 393 Nuclear exportin 1 inhibitor regulates intestinal stem cell fate independently of known differentiation cues and significantly increases paneth cell abundance in organoids.…”