Robust differentiation of human enteroendocrine cells from intestinal stem cells

Abstract

Enteroendocrine (EE) cells are the most abundant hormone-producing cells in humans and are critical regulators of energy homeostasis and gastrointestinal function. Challenges in converting human intestinal stem cells (ISCs) into functional EE cells, ex vivo, have limited progress in elucidating their role in disease pathogenesis and in harnessing their therapeutic potential. To address this, we employed small molecule targeting of the endocannabinoid receptor signaling pathway, JNK, and FOXO1, known to mediate endodermal development and/or hormone production, together with directed differentiation of human ISCs from the duodenum and rectum. We observed marked induction of EE cell differentiation and gut-derived expression and secretion of SST, 5HT, GIP, CCK, GLP-1 and PYY upon treatment with various combinations of three small molecules: rimonabant, SP600125 and AS1842856. Robust differentiation strategies capable of driving human EE cell differentiation is a critical step towards understanding these essential cells and the development of cell-based therapeutics.

Identification of TFPI as a receptor reveals recombination-driven receptor switching in Clostridioides difficile toxin B variants

“…Intestinal organoids are derived from intestinal stem cells and form a single layer of intestinal epithelial cells imbedded in extracellular matrix. Analysis of undifferentiated human enteroids (derived from small intestine), differentiated human rectoids (a proxy for colonoids) 68 , and mouse intestinal organoids revealed each was susceptible to TcdB4.2, based on induced shrinkage, structural disruption, and eventual organoid death quantified by MTT assays (Fig. 4a, b and Supplementary Fig.…”

Pharmacological conversion of gut epithelial cells into insulin-producing cells lowers glycemia in diabetic animals

“…TGFβ, WNT, FGF, Notch, BMP, and FOXO1, along with relevant receptors and signaling pathways, are involved in pancreatic and intestinal tissue patterning (40,41). FOXO1 and Notch signaling interact in determining intestinal stem cell differentiation into Paneth/goblet (42) and EEC lineages (19,21,43). Thus, we combined genetic FoxO1 knockout with pharmacological Notch inhibition (DBZ) to show that dual Notch/FOXO1 inhibition expands the Neurog3 + progenitor pool and its secretory lineage cell descendants.…”

Triple Notch/Tgfβ/FoxO1 blockade converts multiple intestinal sub-lineages into β-like cells and lowers glycemia in diabetic animals

“…Transforming Growth Factor-β (TGF-β), Wnt, fibroblast growth factor (FGF), Notch, bone morphogenic protein (BMP), and FoxO1, along with relevant receptors and signaling pathways, are involved in pancreatic and intestinal tissue patterning (Boonekamp et al, 2020; Nostro et al, 2011). FoxO1 and Notch signaling interact in determining intestinal stem cell differentiation into Paneth/goblet (Ludikhuize et al, 2020) and EEC lineages (Kitamoto et al , 2021; Zeve et al, 2022). Thus, we combined genetic FoxO1 knockout with pharmacological Notch inhibition (DBZ) to show that dual Notch/FoxO1 inhibition expands the Neurog3+ progenitor pool and its secretory lineage cell descendants.…”