Light-stimulated insulin secretion from pancreatic islet-like organoids derived from human pluripotent stem cells

Abstract

Optogenetic techniques permit non-invasive, spatiotemporal, and reversible modulation of cellular activities. Here, we report a novel optogenetic regulatory system for insulin secretion human pluripotent stem cell (hPSC)-derived pancreatic islet like organoids using monSTIM1 (monster-opto-Stromal interaction molecule 1), an ultra-light-sensitive OptoSTIM1 variant. The monSTIM1 transgene was incorporated at AAVS1 locus in human embryonic stem cells (hESCs) CRISPR-Cas9-mediated genome editing. Not only were able to elicit light-induced intracellular Ca2+ concentration ([Ca2+]i) transients from the resulting homozygous monSTIM1+/+-hESCs, but we also successfully differentiated them into pancreatic islet-like organoids (PIOs). Upon light stimulation, the b-cells in these monSTIM1+/+-PIOs displayed reversible and reproducible [Ca2+]i transient dynamics. Furthermore, in response to photoexcitation, they secreted man insulin. Light-responsive insulin secretion was similarly observed in monSTIM1+/+-PIOs produced from neonatal betes (ND) patient-derived induced pluripotent stem cells (iPSCs). Under LED illumination, monSTIM1+/+-PIO-trans-planted diabetic mice produced human c-peptide. Collectively, we developed a cellular model for the optogenetic control of sulin secretion using hPSCs, with the potential to be applied the amelioration of hyperglycemic disorders.