The role of CYP3A4 in amiodarone-associated toxicity on HepG2 cells

Zahno, Anja; Brecht, Karin; Morand, Réjane; Maseneni, Swarna; Török, Michael; Lindinger, Peter W.; Krähenbühl, Stephan

doi:10.1016/j.bcp.2010.11.002

Cited by 94 publications

(69 citation statements)

References 30 publications

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“…Since the cell line does not metabolize amiodarone, the lung macrophages were doped with desethylamiodarone. 37,39 The metabolite has also been shown to accumulate in lung macrophages and contribute to the toxic effect of the drug. 40 The SIMS image shows the protonated molecule of the metabolite, [(C 23 H 25 I 2 NO 3 )+H] + , which was detected at m/z 618.0 (see Figure 4).…”

Section: Experimental Methodsmentioning

confidence: 99%

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Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

et al. 2015

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Section: Experimental Methodsmentioning

confidence: 99%

“…Amiodarone is metabolized by the cytochrome P450 (CYP450) enzymes, specifically cytochrome P450 isoform CYP3A4 37 and CYP2C8 38 in the liver. Since the cell line does not metabolize amiodarone, the lung macrophages were doped with desethylamiodarone.…”

Section: Experimental Methodsmentioning

confidence: 99%

Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

et al. 2015

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“…However, most hepatoma cell lines express low levels of drug-metabolizing enzymes, particularly CYP enzymes, hampering their use in drug metabolism studies. To confer metabolic competence to those cell lines, new cell models that transiently [15][16][17][26][27][28] or stably [18] overexpress heterologous CYP enzymes have been developed and successfully used in toxicity studies. Of these models, HepG2 is the most widely used host cell line for transfection/transduction, since it retains many liver-specific biosynthetic functions, including those for metabolizing activities [29].…”

Section: Resultsmentioning

confidence: 99%

“…As a strategy to overcome this limitation, genetically modified hepatic cell lines expressing human drug metabolizing genes have been developed and used for assessing drug metabolism and toxicity. For example, using adenoviral or lentiviral infection systems, cells that transiently or stably express individual CYPs, such as CYP1A1, CYP2C8, CYP2C9 or CYP3A4, have been generated [15][16][17][18]. These cells responded appropriately to known toxic chemicals, demonstrating their values for toxicity testing and mechanistic studies.…”

Section: Introductionmentioning

confidence: 99%

Development of HepG2-derived cells expressing cytochrome P450s for assessing metabolism-associated drug-induced liver toxicity

Xuan

Chen

Ning

et al. 2016

Chemico-Biological Interactions

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The generation of reactive metabolites from therapeutic agents is one of the major mechanisms of drug-induced liver injury (DILI). In order to evaluate metabolism-related toxicity and improve drug efficacy and safety, we generated a battery of HepG2-derived cell lines that express 14 cytochrome P450s (CYPs) (1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, 3A5 and 3A7) individually using a lentiviral expression system. The expression/production of a specific CYP in each cell line was confirmed by an increased abundance of the CYP at both mRNA and protein levels. Moreover, the enzymatic activities of representative CYPs in the corresponding cell lines were also measured. Using our CYP-expressed HepG2 cells, the toxicity of three drugs that could induce DILI (amiodarone, chlorpromazine and primaquine) was assessed, and all of them showed altered (increased or decreased) toxicity compared to the toxicity in drugtreated wild-type HepG2 cells. CYP-mediated drug toxicity examined in our cell system is consistent with previous reports, demonstrating the potential of these cells for assessing metabolism-related drug toxicity. This cell system provides a practical in vitro approach for drug metabolism screening and for early detection of drug toxicity. It is also a surrogate enzyme source for the enzymatic characterization of a particular CYP that contributes to drug-induced liver toxicity.

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“…Acetaminophen is metabolized by the CYP P450 enzymes 2E1, 1A2, and 3A4 to its reactive intermediate N-acetyl p-benzoquinoneimine, (NAP-QI), which is the main metabolite responsible for the toxicity of acetaminophen (Bessems, andVermeulen, 2001, Patten et al, 1993). Amiodarone is extensively metabolized in the liver by CYP 3A4 to its toxic mono-N-desethyl and di-Ndesethylmetabolites (Zahno et al, 2010). Although primary mouse hepatocytes are a robust model with drug metabolizing capacities, they do show a decline of the CYP 1A2, 3A4, and 2E1 expression with increased cultivation time (Mathijs et al, 2009b).…”

Section: Figmentioning

confidence: 99%

Screening for Drug-Induced Hepatotoxicity in Primary Mouse Hepatocytes Using Acetaminophen, Amiodarone, and Cyclosporin A as Model Compounds: An Omics-Guided Approach

Summeren

Renes

Lizarraga

et al. 2013

OMICS: A Journal of Integrative Biology

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Drug-induced hepatotoxicity is a leading cause of attrition for candidate pharmaceuticals in development. New preclinical screening methods are crucial to predict drug toxicity prior to human studies. Of all in vitro hepatotoxicity models, primary human hepatocytes are considered as 'the gold standard.' However, their use is hindered by limited availability and inter-individual variation. These barriers may be overcome by using primary mouse hepatocytes. We used differential in gel electrophoresis (DIGE) to study large-scale protein expression of primary mouse hepatocytes. These hepatocytes were exposed to three well-defined hepatotoxicants: acetaminophen, amiodarone, and cyclosporin A. Each hepatotoxicant induces a different hepatotoxic phenotype. Based on the DIGE results, the mRNA expression levels of deregulated proteins from cyclosporin A-treated cells were also analyzed. We were able to distinguish cyclosporin A from controls, as well as acetaminophen and amiodarone-treated samples. Cyclosporin A induced endoplasmic reticulum (ER) stress and altered the ER-Golgi transport. Moreover, liver carboxylesterase and bile salt sulfotransferase were differentially expressed. These proteins were associated with a protective adaptive response against cyclosporin A-induced cholestasis. The results of this study are comparable with effects in HepG2 cells. Therefore, we suggest both models can be used to analyze the cholestatic properties of cyclosporin A. Furthermore, this study showed a conserved response between primary mouse hepatocytes and HepG2 cells. These findings collectively lend support for use of omics strategies in preclinical toxicology, and might inform future efforts to better link preclinical and clinical research in rational drug development.

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The role of CYP3A4 in amiodarone-associated toxicity on HepG2 cells

Cited by 94 publications

References 30 publications

Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

Development of HepG2-derived cells expressing cytochrome P450s for assessing metabolism-associated drug-induced liver toxicity

Screening for Drug-Induced Hepatotoxicity in Primary Mouse Hepatocytes Using Acetaminophen, Amiodarone, and Cyclosporin A as Model Compounds: An Omics-Guided Approach

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