Mapping the cellular response to electron transport chain inhibitors reveals selective signaling networks triggered by mitochondrial perturbation
Autor: | Paul Jennings, Giada Carta, Johannes P Schimming, Erik H.J. Danen, Marcel Leist, Giulia Callegaro, Nanette G Vrijenhoek, Joost B. Beltman, Anna Forsby, Johannes Delp, Andrew White, Bob van de Water, Sylvia E. Le Dévédec, Huan Yang, Wanda van der Stel, Salihanur Darici |
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Přispěvatelé: | Molecular and Computational Toxicology, AIMMS |
Jazyk: | angličtina |
Rok vydání: | 2022 |
Předmět: |
Health
Toxicology and Mutagenesis Mitochondrial toxicity ETC complex inhibitors High-content imaging TempO-Seq DILI Oxidative phosphorylation Toxicology ETC Complex Inhibitors Electron Transport ddc:570 medicine Gene silencing Humans Glycolysis Viability assay Electron Transport Complex I Mitochondrial toxicity Chemistry Endoplasmic reticulum Mitochondrial Toxicity ETC complex inhibitors General Medicine Hep G2 Cells medicine.disease Cell biology Mitochondria In Vitro Systems medicine.anatomical_structure High-Content Imaging Hepatocyte Unfolded protein response Hepatocytes TempO-Seq DILI High-content imaging |
Zdroj: | van der Stel, W, Yang, H, Vrijenhoek, N G, Schimming, J P, Callegaro, G, Carta, G, Darici, S, Delp, J, Forsby, A, White, A, le Dévédec, S, Leist, M, Jennings, P, Beltman, J B, van de Water, B & Danen, E H J 2022, ' Mapping the cellular response to electron transport chain inhibitors reveals selective signaling networks triggered by mitochondrial perturbation ', Archives of Toxicology, vol. 96, no. 1, pp. 259-285 . https://doi.org/10.1007/s00204-021-03160-7 Archives of Toxicology, 96 Archives of Toxicology Archives of Toxicology, 96(1), 259-285. Springer Verlag |
ISSN: | 0340-5761 |
Popis: | Mitochondrial perturbation is a key event in chemical-induced organ toxicities that is incompletely understood. Here, we studied how electron transport chain (ETC) complex I, II, or III (CI, CII and CIII) inhibitors affect mitochondrial functionality, stress response activation, and cell viability using a combination of high-content imaging and TempO-Seq in HepG2 hepatocyte cells. CI and CIII inhibitors perturbed mitochondrial membrane potential (MMP) and mitochondrial and cellular ATP levels in a concentration- and time-dependent fashion and, under conditions preventing a switch to glycolysis attenuated cell viability, whereas CII inhibitors had no effect. TempO-Seq analysis of changes in mRNA expression pointed to a shared cellular response to CI and CIII inhibition. First, to define specific ETC inhibition responses, a gene set responsive toward ETC inhibition (and not to genotoxic, oxidative, or endoplasmic reticulum stress) was identified using targeted TempO-Seq in HepG2. Silencing of one of these genes, NOS3, exacerbated the impact of CI and CIII inhibitors on cell viability, indicating its functional implication in cellular responses to mitochondrial stress. Then by monitoring dynamic responses to ETC inhibition using a HepG2 GFP reporter panel for different classes of stress response pathways and applying pathway and gene network analysis to TempO-Seq data, we looked for downstream cellular events of ETC inhibition and identified the amino acid response (AAR) as being triggered in HepG2 by ETC inhibition. Through in silico approaches we provide evidence indicating that a similar AAR is associated with exposure to mitochondrial toxicants in primary human hepatocytes. Altogether, we (i) unravel quantitative, time- and concentration-resolved cellular responses to mitochondrial perturbation, (ii) identify a gene set associated with adaptation to exposure to active ETC inhibitors, and (iii) show that ER stress and an AAR accompany ETC inhibition in HepG2 and primary hepatocytes. Supplementary Information The online version contains supplementary material available at 10.1007/s00204-021-03160-7. |
Databáze: | OpenAIRE |
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