Deriving functional human enteroendocrine cells from pluripotent stem cells

Abstract

Enteroendocrine cells (EECs) are a minor cell population in the intestine yet they play a major role in digestion, satiety and nutrient homeostasis. Recently developed human intestinal organoid models include EECs, but their rarity makes it difficult to study their formation and function. Here, we used the EEC-inducing property of the transcription factor NEUROG3 in human pluripotent stem cell-derived human intestinal organoids and colonic organoids to promote EEC development An 8-h pulse of NEUROG3 expression induced expression of known target transcription factors and after 7 days organoids contained up to 25% EECs in the epithelium. EECs expressed a broad array of human hormones at the mRNA and/or protein level, including motilin, somatostatin, neurotensin, secretin, substance P, serotonin, vasoactive intestinal peptide, oxyntomodulin, GLP-1 and INSL5. EECs secreted several hormones including gastric inhibitory polypeptide (GIP), ghrelin, GLP-1 and oxyntomodulin. Injection of glucose into the lumen of organoids caused an increase in both GIP secretion and K-cell number. Lastly, we observed formation of all known small intestinal EEC subtypes following transplantation and growth of human intestinal organoids in mice.

Intestinal enteroids recapitulate the effects of short-chain fatty acids on the intestinal epithelium

“…Ghrelin which is a hungerinducing hormone, is largely found in the stomach, however it also has been shown to be secreted from enteroendocrine cells in the duodenum [37]. Ghrelin has previously been detected in human intestinal enteroid models [38]. Interestingly, a previous study showed that BUT stimulated ghrelin expression in human gastric-cancer cells [39].…”

ISX-9 manipulates endocrine progenitor fate revealing conserved intestinal lineages in mouse and human

“…Comparatively little is known about the networks controlling endocrine cell fate in the human gut epithelium, but they are assumed to be conserved to some degree, stemming largely from the study of patients with mutations in NGN3 who exhibit intractable malabsorptive diarrhoea due to loss of EECs and a handful of recent studies in organoids (German-Diaz et al, 2017; Pinney et al, 2011; Rubio-Cabezas et al, 2011). In these in vitro studies, a pulse of transgenic NGN3 in iPSCs-derived human organoids mostly recapitulated the predicted endocrine repertoire (Sinagoga et al, 2018). The same transgenic construct has been deployed in organoids derived from tissue resident stem cells and similarly promoted EEC differentiation (Chang-Graham et al, 2019).…”

“…It now appears classical TFs are more promiscuous than lineage tracing implied. Furthermore, there is a paucity of knowledge regarding EEC specification in human intestinal epithelium due to lack of tractable model systems, although, several of the classical TFs are upregulated in response to a NGN3 pulse in intestinal organoids derived from human pluripotent stem cells (Sinagoga et al, 2018; Spence et al, 2011). Understanding the factors which control gut endocrine pedigree has implications for several clinical conditions including diabetes, obesity, gut inflammatory disorders and perhaps cognitive disorders including depression and anxiety.…”

Comparison of human and mouse fetal intestinal tissues reveals differential maturation timelines

“…The early emergence of EECs during human intestinal development raises questions as to why EECs are typically absent in intestinal tissues differentiated from hPSCs in vitro (Fordham et al, 2013;Forster et al, 2014;Nadkarni et al, 2017), when other cells types that arise later in development can be found. The lack of EECs could be attributed to deficiencies in exogenous signaling that activate genes responsible for promoting EEC differentiation, such as Neurogenin-3 (Sinagoga et al, 2018;Spence et al, 2011).…”