Original Abstract

OBJECTIVES: Obesity and type 2 diabetes (T2D) are associated with altered secretion of enteroendocrine hormones, including the glucagon-like peptide-1 (GLP-1). The mechanisms underlying this dysregulation remain poorly understood, partly due to the rarity of enteroendocrine cells (EECs) and technical difficulties in humans. Our aim was to generate an in vitro human jejunal enteroids (HJEs) derived from individuals with obesity and T2D as a simplified intestinal epithelium in a controlled environment, in order to explore GLP-1 secretion in metabolic diseases. SUBJECTS AND METHODS: HJEs were obtained from jejunum fragments sampled during gastric bypass surgery in individuals with severe obesity and normoglycemia (Ob, n = 12), prediabetes (ObPreD, n = 12), or T2D (ObD, n = 10). HJEs were characterized through gene and protein expression analyses and immunofluorescence. To promote the EEC lineage, HJEs were treated with the Notch pathway inhibitor DAPT. Active GLP-1 secretion was assessed by ELISA. RESULTS: HJEs were successfully generated whatever the metabolic group Ob, ObPreD, ObD, exhibiting epithelial cell lineages and expressing critical genes involved in GLP-1 cell lineage, biosynthesis, and secretion. HJEs secreted active GLP-1 in response to both low and high glucose concentrations, regardless of subject metabolic status. However, HJEs from individuals with severe obesity and T2D exhibited a reduced capacity to release GLP-1 in response to high glucose concentrations compared to those from individuals with severe obesity or with obesity and prediabetes. CONCLUSIONS: HJEs represent a robust human-derived model to investigate EEC function and GLP-1 secretion in metabolic diseases. While HJEs retain functional GLP-1-producing cells, their secretory capacity is impaired in the presence of T2D, confirming functional endocrine alterations. These findings support the use of HJEs in preclinical studies targeting enteroendocrine dysfunction.