光遗传学技术允许细胞活动的非侵入性、时空和可逆调节。在这里，我们报道了一种新型光遗传学调节系统，用于使用monSTIM1（monster-opto-Stromal interaction molecule 1）人类多能干细胞（hPSC）衍生的胰岛样器官。通过CRISPR-Cas9介导的基因组编辑，在AAVS1位点中将monSTIM1转基因整合到人类胚胎干细胞（hESCs）中。我们不仅能够从所产生的纯合子monSTIM1+/+-hESCs中引发光诱导的细胞内Ca2+浓度（[Ca2+]i）瞬变，而且还成功将其分化为胰岛样器官（PIOs）。在光刺激下，这些monSTIM1+/+-PIO中的β细胞显示出可逆且可重复的[Ca2+]i瞬变动力学。此外，它们还能分泌人类胰岛素。在新生儿糖尿病（ND）患者来源的诱导多能干细胞（iPSCs）中也观察到类似的光响应胰岛素分泌。在LED照明下，monSTIM1+/+-PIO移植糖尿病小鼠产生人类C肽。总之，我们开发了一种使用hPSCs进行光遗传学控制胰岛素分泌的细胞模型，具有治疗高血糖性疾病的潜力。 版权所有©2023年美国基因和细胞疗法学会。由Elsevier出版。保留所有权利。

Optogenetic techniques permit non-invasive, spatiotemporal, and reversible modulation of cellular activities. Here, we report a novel optogenetic regulatory system for insulin secretion in human pluripotent stem cell (hPSC)-derived pancreatic islet-like organoids using monSTIM1 (monster-opto-Stromal interaction molecule 1). The monSTIM1 transgene was incorporated at the AAVS1 locus in human embryonic stem cells (hESCs) by CRISPR-Cas9-mediated genome editing. Not only were we 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 β-cells in these monSTIM1+/+-PIOs displayed reversible and reproducible [Ca2+]i transient dynamics. Furthermore, in response to photoexcitation, they secreted human insulin. Light-responsive insulin secretion was similarly observed in monSTIM1+/+-PIOs produced from neonatal diabetes (ND) patient-derived induced pluripotent stem cells (iPSCs). Under LED illumination, monSTIM1+/+-PIO-transplanted diabetic mice produced human c-peptide. Collectively, we developed a cellular model for the optogenetic control of insulin secretion using hPSCs, with the potential to be applied to the amelioration of hyperglycemic disorders.Copyright © 2023 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.