Glucose Response by Stem Cell-Derived β Cells In Vitro Is Inhibited by a Bottleneck in Glycolysis

Stem cell-derived β (SC-β) cells could provide unlimited human β cells toward a curative diabetes treatment. Differentiation of SC-β cells yields transplantable islets that secrete insulin in response to glucose challenges. Following transplantation into mice, SC-β cell function is comparable to human islets, but the magnitude and consistency of response in vitro are less robust than observed in cadaveric islets. Here, we profile metabolism of SC-β cells and islets to quantify their capacity to sense glucose and identify reduced anaplerotic cycling in the mitochondria as the cause of reduced glucose-stimulated insulin secretion in SC-β cells. This activity can be rescued by challenging SC-β cells with intermediate metabolites from the TCA cycle and late but not early glycolysis, downstream of the enzymes glyceraldehyde 3-phosphate dehydrogenase and phosphoglycerate kinase. Bypassing this metabolic bottleneck results in a robust, bi-phasic insulin release in vitro that is identical in magnitude to functionally mature human islets. [Display omitted] •Glucose-stimulated insulin secretion (GSIS) in SC-β cells is deficient in vitro•This deficiency is caused by a lack of anaplerotic cycling in high glucose•Metabolites from late glycolysis rescue anaplerotic cycling and insulin secretion•This bottleneck correlates with reduced GAPDH activity in SC-β cells in vitro Glucose-stimulated insulin secretion is deficient in stem cell-derived β (SC-β) cells in vitro. Davis et al. use metabolomic analysis to define a glycolytic bottleneck inhibiting glucose metabolism and sensing in SC-β cells. Cell-permeable intermediates bypass this bottleneck, as does transplantation in vivo, producing insulin secretion indistinguishable from human islets.