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

Choi, Jieun; Shin, Eunji; Lee, Jinsu; Somayadineshraj, D.; Kim, Dongkyu; Shin, Jennifer Hyunjong; Choi, Jin‐Ho; Heo, Won Do; Han, Yong‐Mahn

doi:10.1016/j.ymthe.2023.03.013

Cited by 13 publications

(7 citation statements)

References 66 publications

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“…Recently, the secretion of insulin was reported in pancreatic islet-like organoids of hPSCs carrying a variant of opto-stromal interaction molecule 1 that allows the modulation of intracellular Ca 2+ with light. 23 It should be noted that the elevation of Ca 2+ causes the release of hormone in β-cells irrespective of the concentration of extracellular glucose.…”

Section: ■ Discussionmentioning

confidence: 99%

“…Moreover, the use of human EndoC-βH3 β-cells is clinically relevant, and given their native capacity for glucose sensing and insulin secretion, it is advantageous compared to that of nonpancreatic cells and fusion-derived cells. Recently, the secretion of insulin was reported in pancreatic islet-like organoids of hPSCs carrying a variant of opto-stromal interaction molecule 1 that allows the modulation of intracellular Ca 2+ with light . It should be noted that the elevation of Ca 2+ causes the release of hormone in β-cells irrespective of the concentration of extracellular glucose.…”

Section: Discussionmentioning

confidence: 99%

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Light-Mediated Enhancement of Glucose-Stimulated Insulin Release of Optogenetically Engineered Human Pancreatic Beta-Cells

Chen,

Stoukides,

Tzanakakis

2024

ACS Synth. Biol.

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Enhancement of glucose-stimulated insulin secretion (GSIS) in exogenously delivered pancreatic β-cells is desirable, for example, to overcome the insulin resistance manifested in type 2 diabetes or to reduce the number of β-cells for supporting homeostasis of blood sugar in type 1 diabetes. Optogenetically engineered cells can potentiate their function with exposure to light. Given that cyclic adenosine monophosphate (cAMP) mediates GSIS, we surmised that optoamplification of GSIS is feasible in human β-cells carrying a photoactivatable adenylyl cyclase (PAC). To this end, human EndoC-βH3 cells were engineered to express a blue-light-activated PAC, and a workflow was established combining the scalable manufacturing of pseudoislets (PIs) with efficient adenoviral transduction, resulting in over 80% of cells carrying PAC. Changes in intracellular cAMP and GSIS were determined with the photoactivation of PAC in vitro as well as after encapsulation and implantation in mice with streptozotocin-induced diabetes. cAMP rapidly rose in β-cells expressing PAC with illumination and quickly declined upon its termination. Light-induced amplification in cAMP was concomitant with a greater than 2-fold GSIS vs β-cells without PAC in elevated glucose. The enhanced GSIS retained its biphasic pattern, and the rate of oxygen consumption remained unchanged. Diabetic mice receiving the engineered β-cell PIs exhibited improved glucose tolerance upon illumination compared to those kept in the dark or not receiving cells. The findings support the use of optogenetics for molecular customization of the β-cells toward better treatments for diabetes without the adverse effects of pharmacological approaches.

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Section: ■ Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Light-Mediated Enhancement of Glucose-Stimulated Insulin Release of Optogenetically Engineered Human Pancreatic Beta-Cells

Chen,

Stoukides,

Tzanakakis

2024

ACS Synth. Biol.

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“…In this study, the optogenetic module monSTIM1 (monster‐opto‐stromal interaction molecule 1) was developed to induce Ca 2+ transients thus to promote insulin secretion. 49 …”

Section: Optogenetics‐based Engineered Cell For Insulin and Glp‐1 Pro...mentioning

confidence: 99%

Optogenetic therapeutic strategies for diabetes mellitus

Deng,

Peng,

Yao

et al. 2024

Journal of Diabetes

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Diabetes mellitus (DM) is a common chronic disease affecting humans globally. It is characterized by abnormally elevated blood glucose levels due to the failure of insulin production or reduction of insulin sensitivity and functionality. Insulin and glucagon‐like peptide (GLP)‐1 replenishment or improvement of insulin resistance are the two major strategies to treat diabetes. Recently, optogenetics that uses genetically encoded light‐sensitive proteins to precisely control cell functions has been regarded as a novel therapeutic strategy for diabetes. Here, we summarize the latest development of optogenetics and its integration with synthetic biology approaches to produce light‐responsive cells for insulin/GLP‐1 production, amelioration of insulin resistance and neuromodulation of insulin secretion. In addition, we introduce the development of cell encapsulation and delivery methods and smart bioelectronic devices for the in vivo application of optogenetics‐based cell therapy in diabetes. The remaining challenges for optogenetics‐based cell therapy in the clinical translational study are also discussed.image

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“…Optogenetic protein clustering has provided tools for direct and dynamic study of such clusters with optical precision in living cells and animals. Such methods have allowed control of receptor tyrosine kinases in normal physiology and in cancer [2][3][4][5], stem cell development [6,7], calcium signaling and insulin secretion [8], biomolecular condensates [9,10], compartmentalization of biosynthetic pathways [11], and mammalian cell death [12], among many other applications.…”

Section: Introductionmentioning

confidence: 99%

Rapid optogenetic clustering of a cytoplasmic BcLOV4 variant

Huang,

Benman,

Dong

et al. 2023

Preprint

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Protein clustering is a powerful form of optogenetic control, yet there is currently only one protein, Cry2, whose light-induced clustering has been harnessed for these purposes. Recently, the photoreceptor BcLOV4 was found to form protein clusters in mammalian cells in response to blue light, although clustering coincided with its translocation to the plasma membrane, potentially constraining its application as an optogenetic clustering module. Herein we identify key amino acids that couple clustering to membrane binding, allowing us to engineer a variant of BcLOV4 that clusters in the cytoplasm and does not associate with the membrane in response to blue light. This variant, BcLOVclust, clustered over many cycles with dramatically faster clustering and de-clustering kinetics compared to Cry2. The magnitude of BcLOVclust clustering could be strengthened by appending an intrinsically disordered region from the fused in sarcoma (FUS) protein, or by optimizing the fluorescent protein to which it was fused. BcLOVclust retained the temperature sensitivity of BcLOV4 such that light induced clustering was transient, and the rate of spontaneous declustering increased with temperature. At low temperatures, BcLOVclust and Cry2 could be multiplexed in the same cells, allowing light control of independent protein condensates. BcLOVclust could also be applied to control signaling proteins and stress granules in mammalian cells. Thus BcLOVclust provides an alternative to Cry2 for optogenetic clustering and a method for multiplexed clustering. While its usage is currently suited for organisms that can be cultured below ~30 C, a deeper understanding of BcLOVclust thermal response will further enable its use at physiological mammalian temperatures.

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

Cited by 13 publications

References 66 publications

Light-Mediated Enhancement of Glucose-Stimulated Insulin Release of Optogenetically Engineered Human Pancreatic Beta-Cells

Light-Mediated Enhancement of Glucose-Stimulated Insulin Release of Optogenetically Engineered Human Pancreatic Beta-Cells

Optogenetic therapeutic strategies for diabetes mellitus

Rapid optogenetic clustering of a cytoplasmic BcLOV4 variant

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