HepaRG Cells as Human-Relevant In Vitro Model to Study the Effects of Inflammatory Stimuli on Cytochrome P450 Isoenzymes

Rubin, Katarina; Janefeldt, Annika; Andersson, Linda C.; Berke, Z; Grime, Ken; Andersson, Tommy

doi:10.1124/dmd.114.059246

Cited by 45 publications

(46 citation statements)

References 41 publications

(51 reference statements)

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“…More recent studies have demonstrated cytokine-dependent downregulation of constitutive CYP expression and show that this occurs at physiologically relevant concentrations of cytokines (Aitken & Morgan, 2007;Dallas et al, 2012;Dickmann, Patel, Rock, Wienkers, & Slatter, 2011Evers et al, 2013). Variations on the hepatocyte model include the use of HepaRG cells, a human hepatoma cell line that can be terminally differentiated (Rubin et al, 2015). These reports emphasize that hepatocyte monocultures do not reflect the complex interactions between multiple cytokines and cell types in vivo and cannot accurately predict responses to cytokines and therapeutic proteins in humans (Dickmann et al, 2011Evers et al, 2013).…”

Section: Primary Human Hepatocytesmentioning

confidence: 95%

Role of Cytochrome P450s in Inflammation

Christmas

2015

Advances in Pharmacology

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Section: Primary Human Hepatocytesmentioning

confidence: 95%

Role of Cytochrome P450s in Inflammation

Christmas

2015

Advances in Pharmacology

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“…Taken together, these data demonstrated highly comparable effects of IL-6 on gene expression in PHH and HepaRG, suggesting HepaRG as a valid model to investigate inflammatory responses. The reader is referred to a recent publication (Rubin et al, 2015) that Fig. 7.…”

Section: Discussionmentioning

confidence: 99%

A Systematic Comparison of the Impact of Inflammatory Signaling on Absorption, Distribution, Metabolism, and Excretion Gene Expression and Activity in Primary Human Hepatocytes and HepaRG Cells

et al. 2014

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Inflammatory processes are associated with compromised metabolism and elimination of drugs in the liver, largely mediated by proinflammatory cytokines, such as interleukin-6. The Hepa-RG cell line is an established surrogate for primary human hepatocytes (PHH) in drug metabolism and toxicity studies. However, the impact of inflammatory signaling on HepaRG cells has not been well characterized. In this study, the response of primary human hepatocytes and HepaRG cells to interleukin (IL)-6 was comparatively analyzed. For this purpose, broad-spectrum gene expression profiling, including acute-phase response genes and a large panel of drug-metabolizing enzyme and transporter (DMET) genes as well as their modifiers and regulators, was conducted in combination with cytochrome P450 (P450) activity measurements. Exposure of PHH and HepaRG cells to IL-6 resulted in highly similar coordinated reduction of DMET mRNA, including major ATP-binding cassette transporters (ABCs), P450s, glutathione S-transferases (GSTs), uridine diphosphate glucuronosyltransferases (UGTs), and solute carriers (SLCs). Enzyme activities of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4 were reduced upon 48-72 hours exposure to IL-6 in PHH and HepaRG. However, although these effects were not significant in PHH due to large interindividual donor variability, the impact on HepaRG was more pronounced and highly significant, thus emphasizing the advantage of HepaRG as a more reproducible model system. Exposure of HepaRG cells to interleukin-1b and tumor necrosis factor a resulted in similar effects on gene expression and enzyme activities. The present study emphasizes the role of proinflammatory cytokines in the regulation of drug metabolism and supports the use of HepaRG in lieu of PHH to minimize subject variability.

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“…To test our hypothesis, we investigated the impact of three previously identified disease-associated miRNAs, miR-21, miR-34a, and miR-130b, on the expression and function of a variety of ADME and other genes. As a hepatic cell model we used the HepaRG cell line, which is widely used as a surrogate for primary human hepatocytes because it retains regulated expression of many P450 and ADME genes and also inflammatory signaling (Rubin et al, 2015). Our data show that at least one of these miRNAs, miR130b, has the potential to contribute to a marked down-regulation of certain P450s and other genes during inflammation.…”

Section: Introductionmentioning

confidence: 99%

Inflammation-Associated MicroRNA-130b Down-Regulates Cytochrome P450 Activities and Directly Targets CYP2C9

et al. 2015

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Expression of genes involved in absorption, distribution, metabolism, and excretion (ADME) of drugs is impaired in pathophysiologic conditions such as cholestasis and inflammation. The mechanisms of ADME gene down-regulation remain unclear. In our previous study, strongly elevated levels of microRNAs (miRNA) miR-21, miR-34a, and miR-130b in cholestatic liver and of miR-21 and miR-130b during inflammation were observed. Using HepaRG cells, which retain many functional characteristics of human hepatocytes, we investigated the potential of these miRNAs to down-regulate ADME genes. Cells were transfected with the corresponding miRNA mimics, chemically modified double-stranded RNAs that mimic endogenous miRNAs, followed by mRNA profiling by quantitative reversetranscription polymerase chain reaction. Activities of six cytochrome P450 enzymes (CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4) were determined with a liquid chromatography with tandem mass spectrometric cocktail assay. Although miR-21 and miR-34a showed few effects, transfection of miR-130b led to significantly lower expression of nuclear receptors constitutive androstane receptor (CAR) and farnesoid X receptor (FXRa), the CYPs 1A1, 1A2, 2A6, 2C8, 2C9, and 2C19, as well as GSTA2. Furthermore, miR-130b negatively affected activity levels of all measured P450s by at least 30%. Reporter gene assays employing the CYP2C9 39-untranslated region (39-UTR) confirmed direct regulation by miR-130b. These data support miR-130b as a potential negative regulator of drug metabolism by directly and/or indirectly affecting the expression of several ADME genes. This may be of relevance in pathophysiologic conditions such as cholestasis and inflammation, which are associated with increased miR-130b expression.

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HepaRG Cells as Human-Relevant In Vitro Model to Study the Effects of Inflammatory Stimuli on Cytochrome P450 Isoenzymes

Cited by 45 publications

References 41 publications

Role of Cytochrome P450s in Inflammation

Role of Cytochrome P450s in Inflammation

A Systematic Comparison of the Impact of Inflammatory Signaling on Absorption, Distribution, Metabolism, and Excretion Gene Expression and Activity in Primary Human Hepatocytes and HepaRG Cells

Inflammation-Associated MicroRNA-130b Down-Regulates Cytochrome P450 Activities and Directly Targets CYP2C9

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