Generation of L Cells in Mouse and Human Small Intestine Organoids

Abstract

Upon a nutrient challenge, L cells produce glucagon-like peptide 1 (GLP-1), a powerful stimulant of insulin release. Strategies to augment endogenous GLP-1 production include promoting L-cell differentiation and increasing L-cell number. Here we present a novel in vitro platform to generate functional L cells from three-dimensional cultures of mouse and human intestinal crypts. We show that short-chain fatty acids selectively increase the number of L cells, resulting in an elevation of GLP-1 release. This is accompanied by the upregulation of transcription factors associated with the endocrine lineage of intestinal stem cell development. Thus, our platform allows us to study and modulate the development of L cells in mouse and human crypts as a potential basis for novel therapeutic strategies in patients with type 2 diabetes.

Targeting development of incretin-producing cells increases insulin secretion

“…This indicates that the amplification of GLP-1 secretion was largely dependent on L cell mass and that DBZ treatment did not impair the glucose responsiveness of L cells. We next tested whether the effect of DBZ can be further amplified by SCFAs, which themselves increase L cell numbers in small intestinal organoids by approximately 2-fold (13). SCFAs increased the number of L cells in both DBZ-treated and control mouse organoids ( Figure 1F).…”

“…Finally, the expression of preproglucagon defines the identity of mature L cells, which constitute only 0.5% of intestinal epithelial cells. We have recently shown that short-chain fatty acids (SCFAs) selectively increase the number of L cells in the intestinal epithelium in vitro, followed by a corresponding increase in GLP-1 secretion (13). SCFAs are likely to act through late endocrine precursors by increasing Neurod1 expression (13).…”

“…We have recently shown that short-chain fatty acids (SCFAs) selectively increase the number of L cells in the intestinal epithelium in vitro, followed by a corresponding increase in GLP-1 secretion (13). SCFAs are likely to act through late endocrine precursors by increasing Neurod1 expression (13). It is currently not clear how a change in the number of L cells relates to basal and stimulated GLP-1 concentrations in (patho-)physiological conditions and how it affects insulin secretion and glucose tolerance.…”

Discovery, characterization, and clinical development of the glucagon-like peptides

“…Transgenic technologies have enabled isolation of primary L cell populations suitable for secretion and electrophysiological analyses (91), and stem cell-derived gut organoids, including those of human origin, herald considerable potential for generation of diverse enteroendocrine cell populations from different sites along the gastrointestinal tract (92). Remarkably, despite the theoretical potential of enhancing L cell secretion for the treatment of metabolic disorders (1), there has been scant progress in clinical development of GLP-1 secretagogues exhibiting sustained efficacy in both preclinical and clinical studies.…”

Directed differentiation protocols for successful human intestinal organoids derived from multiple induced pluripotent stem cell lines

“…High expression of pluripotent markers was evident in E2055A, which showed deviations from ideal ESlike morphology within maintenance cultures. Regenerative medicine approaches may be more appropriate by direct differentiation of crypt stem cell enteroids [32,33] rather than iPSC. Overall, these data can help improve cost-effectiveness, and ease of differentiation from different reprogrammed cell lines, enabling HIO to become more commonly available to research labs in this field.…”