Differential toxicity of heterocyclic aromatic amines and their mixture in metabolically competent HepaRG cells

Dumont, Julie; Jossé, Rozenn; Lambert, Carine; Anthérieu, Sébastien; Hégarat, Ludovic Le; Aninat, Caroline; Robin, Marie-Anne; Guguen‐Guillouzo, Christiane; Guillouzo, André

doi:10.1016/j.taap.2010.03.008

Cited by 48 publications

(23 citation statements)

References 36 publications

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“…The novel hepatic in vitro system, the HepaRG cell, has in recent years emerged as a viable new tool for drug discovery, with some success to date as a model for liver toxicity and induction of drug metabolism (Aninat et al, 2006;Guillouzo et al, 2007;Dumont et al, 2010;McGill et al, 2011). HepaRG cells can be routinely produced with expression levels of cytochrome P450 similar to those found in primary hepatocytes and so have potential for use as a model for prediction of clearance-a vital component of drug discovery and one that currently relies on hepatocytes from human liver donors (an unsatisfactory situation because of limits in supply, quality, and variability as discussed widely in the literature).…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, modulation of the concentration of DMSO during the differentiation process can result in expression of cytochrome P450 and transporters at levels similar to or greater than those in primary human hepatocytes in culture (Aninat et al, 2006;Lübberstedt et al, 2011). Since becoming available, HepaRG cells have been used successfully for assessment of cytochrome P450 induction (McGinnity et al, 2009;Anthérieu et al, 2010) and in toxicology studies (Dumont et al, 2010;Lübberstedt et al, 2011;McGill et al, 2011) and appear to have the potential for further understanding of the interplay between drug metabolism, cell metabolism, and liver function in preclinical toxicological studies for either known drugs and new chemical entities. Kanebratt and Andersson (2008) have demonstrated activities of several major drug-metabolizing cytochrome P450s in cultured HepaRG cells, which were of a magnitude similar to that in primary hepatocytes.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of Cryopreserved HepaRG Cells with Cryopreserved Human Hepatocytes for Prediction of Clearance for 26 Drugs

Zanelli¹,

Caradonna²,

Hallifax³

et al. 2011

Drug Metab Dispos

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ABSTRACT:Prediction of clearance in drug discovery currently relies on human primary hepatocytes, which can vary widely in drug-metabolizing enzyme activity. Potential alternative in vitro models include the HepaRG cell (from immortalized hepatoma cells), which in culture can express drug-metabolizing enzymes to an extent comparable to that of primary hepatocytes. Utility of the HepaRG cell will depend on robust performance, relative to that of primary hepatocytes, in routine high-throughput analysis. In this study, we compared intrinsic clearance (CL int ) in the recently developed cryopreserved HepaRG cell system with CL int in human cryopreserved pooled hepatocytes and with CL int in vivo for 26 cytochrome P450 substrate drugs. There was quantitative agreement between CL int in HepaRG cells and human hepatocytes, which was linear throughout the range of CL int (1-2000 ml ⅐ min ؊1 ⅐ kg ؊1 ) and not dependent on particular cytochrome P450 involvement. Prediction of CL int in HepaRG cells was on average within 2-fold of in vivo CL int (using the well stirred liver model), but average fold error was clearance-dependent with greater underprediction (up to at least 5-fold) for the more highly cleared drugs. Recent reporting of this phenomenon in human hepatocytes was therefore confirmed with the hepatocytes used in this study, and hence the HepaRG cell system appears to share an apparently general tendency of clearance-limited CL int in cell models. This study shows the cryopreserved HepaRG cell system to be quantitatively comparable to human hepatocytes for prediction of clearance of drug cytochrome P450 substrates and to represent a promising alternative in vitro tool.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of Cryopreserved HepaRG Cells with Cryopreserved Human Hepatocytes for Prediction of Clearance for 26 Drugs

Zanelli¹,

Caradonna²,

Hallifax³

et al. 2011

Drug Metab Dispos

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“…To evaluate whether the effects caused by the mixture of endosulfan and methoxychlor were additive, more than additive, or less than additive, it was possible to calculate the expected effect of the mixture under the hypothesis of simple additivity response. The expected value was the sum of the effects observed for each individual compound (Dumont et al, 2010). The Mann-Whitney U test was then used to test whether any observed response was significantly different from the expected response.…”

Section: Methodsmentioning

confidence: 99%

Interactions of Endosulfan and Methoxychlor Involving CYP3A4 and CYP2B6 in Human HepaRG Cells

et al. 2014

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Humans are usually exposed to several pesticides simultaneously; consequently, combined actions between pesticides themselves or between pesticides and other chemicals need to be addressed in the risk assessment. Many pesticides are efficient activators of pregnane X receptor (PXR) and/or constitutive androstane receptor (CAR), two major nuclear receptors that are also activated by other substrates. In the present work, we searched for interactions between endosulfan and methoxychlor, two organochlorine pesticides whose major routes of metabolism involve CAR-and PXR-regulated CYP3A4 and CYP2B6, and whose mechanisms of action in humans remain poorly understood. For this purpose, HepaRG cells were treated with both pesticides separately or in mixture for 24 hours or 2 weeks at concentrations relevant to human exposure levels. In combination they exerted synergistic cytotoxic effects. Whatever the duration of treatment, both compounds increased CYP3A4 and CYP2B6 mRNA levels while differently affecting their corresponding activities. Endosulfan exerted a direct reversible inhibition of CYP3A4 activity that was confirmed in human liver microsomes. By contrast, methoxychlor induced this activity. The effects of the mixture on CYP3A4 activity were equal to the sum of those of each individual compound, suggesting an additive effect of each pesticide. Despite CYP2B6 activity being unchanged and increased with endosulfan and methoxychlor, respectively, no change was observed with their mixture, supporting an antagonistic effect. Altogether, our data suggest that CAR and PXR activators endosulfan and methoxychlor can interact together and with other exogenous substrates in human hepatocytes. Their effects on CYP3A4 and CYP2B6 activities could have important consequences if extrapolated to the in vivo situation.

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“…CYP2E1 activity was subsequently assessed by measuring hydroxylation of aniline into p-aminophenol (Robin et al, 2005b) and hydroxylation of chlorzoxazone into 6-hydroxychlorzoxazone (Guillouzo and Chesne, 1996). Hepatic caspase 3 activity was assessed in liver homogenates as described previously (Dumont et al, 2010).…”

Section: Methodsmentioning

confidence: 99%

Differences in Early Acetaminophen Hepatotoxicity between Obeseob/obanddb/dbMice

Aubert

Begriche

Delannoy

et al. 2012

J Pharmacol Exp Ther

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Clinical investigations suggest that hepatotoxicity after acetaminophen (APAP) overdose could be more severe in the context of obesity and nonalcoholic fatty liver disease. The preexistence of fat accumulation and CYP2E1 induction could be major mechanisms accounting for such hepatic susceptibility. To explore this issue, experiments were performed in obese diabetic ob/ob and db/db mice. Preliminary investigations performed in male and female wild-type, ob/ob, and db/db mice showed a selective increase in hepatic CYP2E1 activity in female db/db mice. However, liver triglycerides in these animals were significantly lower compared with ob/ob mice. Next, APAP (500 mg/kg) was administered in female wild-type, ob/ ob, and db/db mice, and investigations were carried out 0.5, 2, 4, and 8 h after APAP intoxication. Liver injury 8 h after APAP intoxication was higher in db/db mice, as assessed by plasma transaminases, liver histology, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. In db/db mice, however, the extent of hepatic glutathione depletion, levels of APAP-protein adducts, c-Jun N-terminal kinase activation, changes in gene expression, and mitochondrial DNA levels were not greater compared with the other genotypes. Furthermore, in the db/db genotype plasma lactate and ␤-hydroxybutyrate were not specifically altered, whereas the plasma levels of APAP-glucuronide were intermediary between wild-type and ob/ob mice. Thus, early APAP-induced hepatotoxicity was greater in db/db than ob/ob mice, despite less severe fatty liver and similar basal levels of transaminases. Hepatic CYP2E1 induction could have an important pathogenic role when APAP-induced liver injury occurs in the context of obesity and related metabolic disorders.

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Differential toxicity of heterocyclic aromatic amines and their mixture in metabolically competent HepaRG cells

Cited by 48 publications

References 36 publications

Comparison of Cryopreserved HepaRG Cells with Cryopreserved Human Hepatocytes for Prediction of Clearance for 26 Drugs

Comparison of Cryopreserved HepaRG Cells with Cryopreserved Human Hepatocytes for Prediction of Clearance for 26 Drugs

Interactions of Endosulfan and Methoxychlor Involving CYP3A4 and CYP2B6 in Human HepaRG Cells

Differences in Early Acetaminophen Hepatotoxicity between Obeseob/obanddb/dbMice

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