EndoC-βH1 cells display increased sensitivity to sodium palmitate when cultured in DMEM/F12 medium

Krizhanovskii, Camilla; Kristinsson, Hjalti; Elksnis, Andris; Wang, Xuan; Gavali, Hamid; Bergsten, Peter; Scharfmann, Raphaël; Welsh, Nils

doi:10.1080/19382014.2017.1296995

Cited by 28 publications

(27 citation statements)

References 10 publications

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“…For the glucolipotoxic stress, three days of treatment with 0.1–0.7 mM palmitate in the presence of 20 mM glucose did not initiate apoptotic cell death in the EndoC-βH1 cells. Similar results were obtained in a recent study using 1 mM palmitate and 22 mM glucose [42] . Interestingly, in the same study it was found that the use of 1.5–2 mM palmitate resulted in an increase in cell death already at day 1 and that use of the more nutrient rich culture medium DMEM/Ham's F12 significantly increased palmitate induced cell death even at normal glucose level (5.5 mM).…”

Section: Discussionsupporting

confidence: 91%

The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates

Tsonkova

Sand

Wolf

et al. 2018

Molecular Metabolism

117

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ObjectiveTo characterize the EndoC-βH1 cell line as a model for human beta cells and evaluate its beta cell functionality, focusing on insulin secretion, proliferation, apoptosis and ER stress, with the objective to assess its potential as a screening platform for identification of novel anti-diabetic drug candidates.MethodsEndoC-βH1 was transplanted into mice for validation of in vivo functionality. Insulin secretion was evaluated in cells cultured as monolayer and as pseudoislets, as well as in diabetic mice. Cytokine induced apoptosis, glucolipotoxicity, and ER stress responses were assessed. Beta cell relevant mRNA and protein expression were investigated by qPCR and antibody staining. Hundreds of proteins or peptides were tested for their effect on insulin secretion and proliferation.ResultsTransplantation of EndoC-βH1 cells restored normoglycemia in streptozotocin induced diabetic mice. Both in vitro and in vivo, we observed a clear insulin response to glucose, and, in vitro, we found a significant increase in insulin secretion from EndoC-βH1 pseudoislets compared to monolayer cultures for both glucose and incretins.Apoptosis and ER stress were inducible in the cells and caspase 3/7 activity was elevated in response to cytokines, but not affected by the saturated fatty acid palmitate.By screening of various proteins and peptides, we found Bombesin (BB) receptor agonists and Pituitary Adenylate Cyclase-Activating Polypeptides (PACAP) to significantly induce insulin secretion and the proteins SerpinA6, STC1, and APOH to significantly stimulate proliferation.ER stress was readily induced by Tunicamycin and resulted in a reduction of insulin mRNA. Somatostatin (SST) was found to be expressed by 1% of the cells and manipulation of the SST receptors was found to significantly affect insulin secretion.ConclusionsOverall, the EndoC-βH1 cells strongly resemble human islet beta cells in terms of glucose and incretin stimulated insulin secretion capabilities. The cell line has an active cytokine induced caspase 3/7 apoptotic pathway and is responsive to ER stress initiation factors. The cells' ability to proliferate can be further increased by already known compounds as well as by novel peptides and proteins. Based on its robust performance during the functionality assessment assays, the EndoC-βH1 cell line was successfully used as a screening platform for identification of novel anti-diabetic drug candidates.

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Section: Discussionsupporting

confidence: 91%

The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates

Tsonkova

Sand

Wolf

et al. 2018

Molecular Metabolism

117

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“…Rat and mouse beta cells are highly sensitive to palmitate treatment that induces dysfunction and apoptosis [3]. On the other hand, previous data indicated that treatment of EndoC-βH1 cells with palmitate does not induce lipotoxicity under standard culture conditions [51,52]. Our current data further confirm this.…”

Section: Discussionsupporting

confidence: 87%

“…By investigating saturated NEFA metabolism and its related enzymes through knockdown using siRNA, we identified SCD as the main brake on palmitate toxicity. SCD is highly expressed in primary human beta cells ( [50,51] and the present study). Interestingly, elevated SCD levels have been shown to protect against saturated NEFA in a number of cell types, including the mouse beta cell line MIN6 cells and human islets [21,48,49].…”

Section: Discussionmentioning

confidence: 51%

Stearoyl CoA desaturase is a gatekeeper that protects human beta cells against lipotoxicity and maintains their identity

et al. 2019

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Aims/hypothesis During the onset of type 2 diabetes, excessive dietary intake of saturated NEFA and fructose lead to impaired insulin production and secretion by insulin-producing pancreatic beta cells. The majority of data on the deleterious effects of lipids on functional beta cell mass were obtained either in vivo in rodent models or in vitro using rodent islets and beta cell lines. Translating data from rodent to human beta cells remains challenging. Here, we used the human beta cell line EndoC-βH1 and analysed its sensitivity to a lipotoxic and glucolipotoxic (high palmitate with or without high glucose) insult, as a way to model human beta cells in a type 2 diabetes environment. Methods EndoC-βH1 cells were exposed to palmitate after knockdown of genes related to saturated NEFA metabolism. We analysed whether and how palmitate induces apoptosis, stress and inflammation and modulates beta cell identity. Results EndoC-βH1 cells were insensitive to the deleterious effects of saturated NEFA (palmitate and stearate) unless stearoyl CoA desaturase (SCD) was silenced. SCD was abundantly expressed in EndoC-βH1 cells, as well as in human islets and human induced pluripotent stem cell-derived beta cells. SCD silencing induced markers of inflammation and endoplasmic reticulum stress and also IAPP mRNA. Treatment with the SCD products oleate or palmitoleate reversed inflammation and endoplasmic reticulum stress. Upon SCD knockdown, palmitate induced expression of dedifferentiation markers such as SOX9, MYC and HES1. Interestingly, SCD knockdown by itself disrupted beta cell identity with a decrease in mature beta cell markers INS, MAFA and SLC30A8 and decreased insulin content and glucose-stimulated insulin secretion. Conclusions/interpretation The present study delineates an important role for SCD in the protection against lipotoxicity and in the maintenance of human beta cell identity. Data availability Microarray data and all experimental details that support the findings of this study have been deposited in in the GEO database with the GSE130208 accession code.

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“…Human islets were cultured in Connaught Medical Research Laboratories (CMRL) 1066 medium containing 5.5 mM glucose supplemented with 10% (vol/vol) FCS, 100 IU/mL penicillin, 100 μg/mL streptomycin, and 2 mM l -glutamine. Human beta-cell line EndoC-βH1, kindly provided by R. Scharfmann and P. Ravassard (INSERM, Paris) ( 40 ), was maintained in DMEM/F12 medium ( 41 ) containing 5.5 mM glucose with 0.2% (vol/vol) MycoZap (Lonza), 10 mM nicotinamide, 6.7 ng/mL selenite, 5.5 µg/mL transferrin, 2% (wt/vol) fatty acid-free albumin, 50 µM β-mercaptoethanol, and 2 mM l -glutamine and cultured on ECM/fibronectin (Sigma)-coated surfaces. Islets and cells were cultured at 37 °C in 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

BRICHOS domain of Bri2 inhibits islet amyloid polypeptide (IAPP) fibril formation and toxicity in human beta cells

Oskarsson

Hermansson

Wang

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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SignificanceAccumulation of islet amyloid polypeptide (IAPP)-containing amyloid fibrils is the main pathological finding in pancreatic islets in type 2 diabetes. The formation of these IAPP amyloid fibrils is considered toxic and may constitute a major cause for the loss of insulin-producing beta cells. The protein domain BRICHOS is present in several different proproteins and possesses antiamyloid chaperone activity. This study demonstrates expression of the BRICHOS-containing protein Bri2 in human pancreatic beta cells and its colocalization with IAPP. The Bri2 BRICHOS domain effectively prevents IAPP from forming fibrils and protects cells from the toxicity associated with IAPP fibrillation. It is concluded that the Bri2 BRICHOS domain may act as an endogenous inhibitor of IAPP amyloid formation in pancreatic beta cells.

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EndoC-βH1 cells display increased sensitivity to sodium palmitate when cultured in DMEM/F12 medium

Cited by 28 publications

References 10 publications

The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates

The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates

Stearoyl CoA desaturase is a gatekeeper that protects human beta cells against lipotoxicity and maintains their identity

BRICHOS domain of Bri2 inhibits islet amyloid polypeptide (IAPP) fibril formation and toxicity in human beta cells

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