한빛사논문
Jieun Choi 1,5, Eunji Shin 1,5, Jinsu Lee 1,5, Somayadineshraj Devarasou 2, Dongkyu Kim 1, Jennifer H. Shin 2, Jin-Ho Choi 3, Won Do Heo 1, Yong-Mahn Han 1,4
1Department of Biological Sciences, KAIST, Daejeon 34141, Republic of Korea
2Department of Mechanical Engineering, KAIST, Daejeon 34141, Republic of Korea
3Department of Pediatrics, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea
4Graduate School of Medical Science and Engineering, KAIST, Daejeon 34141, Republic of Korea
5These authors contributed equally
Corresponding authors: Won Do Heo, Yong-Mahn Han
Abstract
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), an ultra-light-sensitive OptoSTIM1 variant. 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.
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