The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates
| العنوان: | The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates |
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| المؤلفون: | Rasmus Wernersson, Fredrik Wolfhagen Sand, Charlotte Helgstrand, Kevin Dalgaard, Jacob Hald, Anne Grapin-Botton, Sven Hastrup, Thomas Frogne, Jonas Ahnfelt-Rønne, Ole D. Madsen, Lena Hansson, Erik Vernet, Mark Kalisz, Dennis Madsen, Tomas Alanentalo, Mads Grønborg, Maja Borup Kjær Petersen, Fredrik Kryh Öberg, Michael Paolo Bastner Sandrini, Annette Plesner, Claude Rescan, Christian Honoré, Philippe Ravassard, Carsten Jessen, Louise Winkel, Camilla Ingvorsen, Xenia Asbæk Wolf, Hanni Willenbrock, Jan N. Jensen, Anna Kirstine Ringgaard, Christine Bruun, Violeta Georgieva Tsonkova, Jacob Hecksher-Sørensen, Johannes Josef Fels, Lars Groth Grunnet, Allan Christian Shaw |
| المساهمون: | University of Copenhagen = Københavns Universitet (KU), Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | Molecular metabolism Molecular metabolism, Elsevier, 2018, 8, pp.144-157. ⟨10.1016/j.molmet.2017.12.007⟩ Molecular Metabolism Molecular Metabolism, Vol 8, Iss, Pp 144-157 (2018) Tsonkova, V G, Sand, F W, Wolf, X A, Grunnet, L G, Ringgaard, A K, Ingvorsen, C, Winkel, L, Kalisz, M, Dalgaard, K, Bruun, C, Fels, J J, Helgstrand, C, Hastrup, S, Öberg, F K, Vernet, E, Sandrini, M P B, Shaw, A C, Jessen, C, Grønborg, M, Hald, J, Willenbrock, H, Madsen, D, Wernersson, R, Hansson, L, Jensen, J N, Plesner, A, Alanentalo, T, Petersen, M B K, Grapin-Botton, A, Honoré, C, Ahnfelt-Rønne, J, Hecksher-Sørensen, J, Ravassard, P, Madsen, O D, Rescan, C & Frogne, T 2018, ' The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates ', Molecular Metabolism, vol. 8, pp. 144-157 . https://doi.org/10.1016/j.molmet.2017.12.007Test |
| بيانات النشر: | HAL CCSD, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, SI, Stimulation index, medicine.medical_treatment, EndoC-βH1, Proliferation, Cell Culture Techniques, Drug Evaluation, Preclinical, Mice, SCID, CytMix, Cytokine mixture, Mice, 0302 clinical medicine, Insulin-Secreting Cells, Insulin Secretion, Cells, Cultured, Chemistry, Bxv1, B10 xenotropic virus 1, EdU, 5-ethynyl-2′-deoxyuridine, 3. Good health, Cell biology, Cytokine, Original Article, Beta cell, Glucose stimulated insulin secretion, medicine.drug, lcsh:Internal medicine, Somatostatin signaling, Incretin, GSIS, Glucose stimulated insulin secretion, 030209 endocrinology & metabolism, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Cell Line, Diabetes Mellitus, Experimental, 03 medical and health sciences, GLP1R, GLP1 receptor, PACAP, Pituitary Adenylate Cyclase-Activating Polypeptides, medicine, Animals, Humans, Hypoglycemic Agents, Pseudoislets, lcsh:RC31-1245, sXBP1, spliced XBP1, Molecular Biology, IPGTT, Intraperitoneal glucose tolerance test, Insulin, SST, Somatostatin, BB, Bombesin, STZ, Streptozotocin, Cell Biology, Streptozotocin, SSTR, Somatostatin receptor, SEM, Standard error of the mean, 030104 developmental biology, Cell culture, Apoptosis, Unfolded protein response, Ex4, Exendin-4 |
| الوصف: | Objective To 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. Methods EndoC-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. Results Transplantation 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. Conclusions Overall, 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. Highlights • The EndoC-βH1 cell line responds robustly to glucose and incretins and restores normoglycaemia in diabetic mice. • EndoC-βH1 cells assembled in pseudoislets show an improved insulin secretion profile compared with monolayer cells. • Bombesin receptor agonists as well as the neuropeptides PACAP27 and PACAP38 significantly increase insulin secretion. • The proteins SerpineA6, STC1 and APOH significantly increase the rate of proliferation. • EndoC-βH1 cells are affected in a biological relevant way by cytokines and are sensitive to Tunicamycin induced ER stress. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 2212-8778 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5cb3e641ee305f9db5f81e09f4ed88e7Test https://hal.sorbonne-universite.fr/hal-02018209/documentTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5cb3e641ee305f9db5f81e09f4ed88e7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 22128778 |
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