The utility of a differentiated preclinical liver model, HepaRG cells, in investigating delayed toxicity via inhibition of mitochondrial-replication induced by fialuridine
| العنوان: | The utility of a differentiated preclinical liver model, HepaRG cells, in investigating delayed toxicity via inhibition of mitochondrial-replication induced by fialuridine |
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| المؤلفون: | Carol E. Jolly, Rosalind E. Jenkins, B. Kevin Park, Alison J. Beckett, Jan Snoeys, Amy E. Chadwick, Sophie L. Penman, Oisin Douglas, Laleh Kamalian, Mario Monshouwer, Dominic P. Williams, Damir Simic |
| المصدر: | Toxicology and Applied Pharmacology |
| بيانات النشر: | Elsevier BV, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Mitochondrion, Pharmacology, Toxicology, 0302 clinical medicine, CE, coupling efficiency, Cytotoxic T cell, PHH, primary human hepatocytes, ALR, ATP-linked respiration, mtDNA, Chemistry, SRC, spare respiratory capacity, PL, proton leak, Mitochondria, DILI, drug-induced liver injury, Mitochondrial toxicity, Liver, hENT1, human equilibrative nucleoside transporter 1, MR, maximal respiration, 030220 oncology & carcinogenesis, Toxicity, HepaRG, medicine.drug, DNA Replication, Mitochondrial DNA, Fialuridine, Antiviral Agents, DNA, Mitochondrial, Thymidylate kinase, Article, 03 medical and health sciences, FIAM, pharmacologically inactive epimer of fialuridine, TK1, TK2, thymidine kinase-1 and -2, Cell Line, Tumor, medicine, Humans, TEM, transmission electron microscopy, Cmax, maximal plasma concentration, Dose-Response Relationship, Drug, Hepatotoxicity, Arabinofuranosyluracil, BR, basal respiration, medicine.disease, In vitro, mtDNA, mitochondrial DNA, 030104 developmental biology, FIAU, fialuridine, Hepatocytes |
| الوصف: | During its clinical development fialuridine caused liver toxicity and the death of five patients. This case remains relevant due to the continued development of mechanistically-related compounds against a back-drop of simple in vitro models which remain limited for the preclinical detection of such delayed toxicity. Here, proteomic investigation of a differentiated, HepaRG, and proliferating, HepG2 cell model was utilised to confirm the presence of the hENT1 transporter, thymidine kinase-1 and -2 (TK1, TK2) and thymidylate kinase, all essential in order to reproduce the cellular activation and disposition of fialuridine in the clinic. Acute metabolic modification assays could only identify mitochondrial toxicity in HepaRG cells following extended dosing, 2 weeks. Toxic effects were observed around 10 μM, which is within a range of 10–15 X approximate Cmax. HepaRG cell death was accompanied by a significant decrease in mitochondrial DNA content, indicative of inhibition of mitochondrial replication, and a subsequent reduction in mitochondrial respiration and the activity of mitochondrial respiratory complexes, not replicated in HepG2 cells. The structural epimer of fialuridine, included as a pharmacological negative control, was shown to have no cytotoxic effects in HepaRG cells up to 4 weeks. Overall, these comparative studies demonstrate the HepaRG model has translational relevance for fialuridine toxicity and therefore may have potential in investigating the inhibition of mitochondrial replication over prolonged exposure for other toxicants. Highlights • Metabolic modification has been adapted to identify delayed (weeks) mitochondrial dysfunction. • Fialuridine induces delayed mitochondrial dysfunction and toxicity at physiologically relevant exposures. • Inhibition of mitochondrial replication leads to decreased oxidative phosphorylation before cell death. • HepaRG cells are a more suitable model for fialuridine-induced toxicity than HepG2 cells. • HepaRG cells offer a practical model to study toxicity via effects on mitochondrial replication. |
| تدمد: | 0041-008X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bc33f6f2025f2dc67b6c695ac4dfd787 https://doi.org/10.1016/j.taap.2020.115163 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....bc33f6f2025f2dc67b6c695ac4dfd787 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 0041008X |
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