Modeling Congenital Hyperinsulinism with ABCC8-Deficient Human Embryonic Stem Cells Generated by CRISPR/Cas9

Guo, Dongsheng; Liu, Haikun; Ruzi, Aynisahan; Gao, Ge; Abbas, Nasir; Li, Yanli; Yang, Fan; Wu, Fa-Quan; Xu, Guosheng; Li, Yin-xiong

doi:10.1038/s41598-017-03349-w

Cited by 18 publications

(14 citation statements)

References 54 publications

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“…CRISPR-Cas9 genome editing [18] has made it possible to efficiently correct specific mutations in order to generate isogenic cell lines that are discordant only for the mutation of interest [19,20]. Differentiating human embryonic stem cells with a K ATP HI mutation in 2D towards beta-like cells has been previously shown to replicate the insulin hypersecretion phenotype in vitro [21].…”

Section: Introductionmentioning

confidence: 99%

SUR1-mutant iPS cell-derived islets recapitulate the pathophysiology of congenital hyperinsulinism

et al. 2021

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Aims/hypothesis Congenital hyperinsulinism caused by mutations in the K ATP-channel-encoding genes (K ATP HI) is a potentially life-threatening disorder of the pancreatic beta cells. No optimal medical treatment is available for patients with diazoxideunresponsive diffuse K ATP HI. Therefore, we aimed to create a model of K ATP HI using patient induced pluripotent stem cell (iPSC)-derived islets. Methods We derived iPSCs from a patient carrying a homozygous ABCC8 V187D mutation, which inactivates the sulfonylurea receptor 1 (SUR1) subunit of the K ATP-channel. CRISPR-Cas9 mutation-corrected iPSCs were used as controls. Both were differentiated to stem cell-derived islet-like clusters (SC-islets) and implanted into NOD-SCID gamma mice. Results SUR1-mutant and-corrected iPSC lines both differentiated towards the endocrine lineage, but SUR1-mutant stem cells generated 32% more beta-like cells (SC-beta cells) (64.6% vs 49.0%, p = 0.02) and 26% fewer alpha-like cells (16.1% vs 21.8% p = 0.01). SUR1-mutant SC-beta cells were 61% more proliferative (1.23% vs 0.76%, p = 0.006), and this phenotype could be induced in SUR1-corrected cells with pharmacological K ATP-channel inactivation. The SUR1-mutant SC-islets secreted 3.2-fold more insulin in low glucose conditions (0.0174% vs 0.0054%/min, p = 0.0021) and did not respond to K ATP-channel-acting drugs in vitro. Mice carrying grafts of SUR1-mutant SC-islets presented with 38% lower fasting blood glucose (4.8 vs 7.7 mmol/l, p = 0.009) and their grafts failed to efficiently shut down insulin secretion during induced hypoglycaemia. Explanted SUR1-mutant grafts displayed an increase in SC-beta cell proportion and SC-beta cell nucleomegaly, which was independent of proliferation. Conclusions/interpretation We have created a model recapitulating the known pathophysiology of K ATP HI both in vitro and in vivo. We have also identified a novel role for K ATP-channel activity during human islet development. This model will enable further studies for the improved understanding and clinical management of K ATP HI without the need for primary patient tissue. Keywords Beta cells. Congenital hyperinsulinism. Disease modelling. Induced pluripotent stem cells. K ATP-channel. Pancreatic islet development. Stem cell-derived islets Abbreviations CHI Congenital hyperinsulinism CXCR4 C-X-C motif chemokine receptor 4 GBC Glibenclamide iPSC Induced pluripotent stem cell K ATP HI Congenital hyperinsulinism due to loss-of-function of the K ATP channel PDX1 Pancreatic and duodenal homeobox 1 RNP Ribonucleoprotein SC-alpha cell Stem cell-derived alpha-like cell * Väinö Lithovius

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Section: Introductionmentioning

confidence: 99%

SUR1-mutant iPS cell-derived islets recapitulate the pathophysiology of congenital hyperinsulinism

et al. 2021

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“…The repression of Kir6.2/SUR1 channels stimulates the release of insulin [60], and it has been reported that mutations and deficiencies of ABCC8 and KCNJ11 induce hyperinsulinism [61][62][63][64][65]. In various studies and clinical reports, the hypersecretion of insulin has been linked to the development of several types of cancer, including breast, colon, liver, and kidney cancers [66,67], and the survival rates through GEPIA in our research also confirmed that higher the expression levels of the two genes, the better is the survival rate ( Figure S4A).…”

Section: Discussionmentioning

confidence: 99%

CeRNA Network Analysis Representing Characteristics of Different Tumor Environments Based on 1p/19q Codeletion in Oligodendrogliomas

Ahn

Park

Kang

et al. 2020

Cancers

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Oligodendroglioma (OD) is a subtype of glioma occurring in the central nervous system. The 1p/19q codeletion is a prognostic marker of OD with an isocitrate dehydrogenase (IDH) mutation and is associated with a clinically favorable overall survival (OS); however, the exact underlying mechanism remains unclear. Long non-coding RNAs (lncRNAs) have recently been suggested to regulate carcinogenesis and prognosis in cancer patients. Here, we performed in silico analyses using low-grade gliomas from datasets obtained from The Cancer Genome Atlas to investigate the effects of ceRNA with 1p/19q codeletion on ODs. Thus, we selected modules of differentially expressed genes that were closely related to 1p/19q codeletion traits using weighted gene co-expression network analysis and constructed 16 coding RNA–miRNA–lncRNA networks. The ceRNA network participated in ion channel activity, insulin secretion, and collagen network and extracellular matrix (ECM) changes. In conclusion, ceRNAs with a 1p/19q codeletion can create different tumor microenvironments via potassium ion channels and ECM composition changes; furthermore, differences in OS may occur. Moreover, if extrapolated to gliomas, our results can provide insights into the consequences of identical gene expression, indicating the possibility of tracking different biological processes in different subtypes of glioma.

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“…Congenital hyperinsulinism (CHI), a rare genetic disorder related to mutations in sulfonylurea receptor 1 (SUR1), encoded by ABCC8 , is characterized by excessive insulin secretion and hypoglycemia (Nessa et al, 2015 ). ABCC8- deficient sc-β cells recapitulated the disease phenotypes of CHI in vitro , indicating that these cells could be an attractive model for further elucidating SUR1 function (Guo et al, 2017 ). We have summarized the disease models related to islet organoids and sc-β cells in Table 1 .…”

Section: Applications Of Islet Organoidsmentioning

confidence: 99%

Islet organoid as a promising model for diabetes

Zhang

Song

et al. 2021

Protein Cell

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Studies on diabetes have long been hampered by a lack of authentic disease models that, ideally, should be unlimited and able to recapitulate the abnormalities involved in the development, structure, and function of human pancreatic islets under pathological conditions. Stem cell-based islet organoids faithfully recapitulate islet development in vitro and provide large amounts of three-dimensional functional islet biomimetic materials with a morphological structure and cellular composition similar to those of native islets. Thus, islet organoids hold great promise for modeling islet development and function, deciphering the mechanisms underlying the onset of diabetes, providing an in vitro human organ model for infection of viruses such as SARS-CoV-2, and contributing to drug screening and autologous islet transplantation. However, the currently established islet organoids are generally immature compared with native islets, and further efforts should be made to improve the heterogeneity and functionality of islet organoids, making it an authentic and informative disease model for diabetes. Here, we review the advances and challenges in the generation of islet organoids, focusing on human pluripotent stem cell-derived islet organoids, and the potential applications of islet organoids as disease models and regenerative therapies for diabetes.

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Modeling Congenital Hyperinsulinism with ABCC8-Deficient Human Embryonic Stem Cells Generated by CRISPR/Cas9

Cited by 18 publications

References 54 publications

SUR1-mutant iPS cell-derived islets recapitulate the pathophysiology of congenital hyperinsulinism

SUR1-mutant iPS cell-derived islets recapitulate the pathophysiology of congenital hyperinsulinism

CeRNA Network Analysis Representing Characteristics of Different Tumor Environments Based on 1p/19q Codeletion in Oligodendrogliomas

Islet organoid as a promising model for diabetes

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