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

Abstract

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.

The protein synthesis inhibitor brusatol normalizes high‐fat diet‐induced glucose intolerance in male C57BL/6 mice: role of translation factor eIF5A hypusination

“…We also analyzed whether brusatol affects eIF5A hypusination in human EndoC-bH1 cells, which are considered to be valid a model for primary human islet cells (30). As in RIN-5AH cells, brusatol and GC7 reduced eIF5A hypusination dose-and time-dependently in EndoC-bH1 cells (Fig.…”

Myt Transcription Factors prevent stress-response gene over-activation to enable postnatal pancreatic β cell proliferation and function

“…To provide further support for MYT TF action in stress-pathway gene regulation in human β cells, siRNA-based MYT knockdown was performed in the human EndoC-βH1 cell line, which displays functional properties similar to primary human islet β cells (Tsonkova et al, 2018). MYT1, 2, and 3 were all detected and MYT siRNA treatment effectively reduced their protein and mRNA levels in a majority of EndoC-βH1 cells (Figure 7).…”

The supply chain of human pancreatic β cell lines

“…All of the above examples depended on experiments that would have been difficult to perform with human islets because of limitations in cell purity or cell number and that would have produced different results in rodent β cell lines because of species differences. Interestingly, human β cell lines have now been validated by the pharmaceutical industry as a screening model to identify novel drug target candidates (87); to determine the effects of fatty acid esters of hydroxylated fatty acids on glucose-stimulated insulin secretion (88); to study the mechanism of secretagogin release by human β cells (89); and to further test a gastrointestinal peptide-based (GIP-based) dual incretin receptor agonist (90).…”