Interindividual Variability in Gene Expression Profiles in Human Hepatocytes and Comparison with HepaRG Cells

Rogue, Alexandra; Lambert, Carine; Spire, Catherine; Claude, Nancy; Guillouzo, André

doi:10.1124/dmd.111.042028

Cited by 58 publications

(51 citation statements)

References 30 publications

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“…Furthermore, SULT1B1 mRNA expression was slightly decreased, but still highly variable. GSTP1 could not be detected in HepaRG cells, as previously shown in the literature (Rogue et al, 2012). Also, in contrast to findings in PHH, the mRNA expression of ABCB1 and ABCC2 was not significantly downregulated, while SLC22A7 was much stronger downregulated (Fig.…”

supporting

confidence: 67%

“…However, PHH are restricted in availability and have a limited life span. Furthermore, PHH exhibit marked interindividual variations, including variability in expression and corresponding activities of many genes related to drug metabolism (Rogue et al, 2012). The HepaRG cell line has been proposed as an alternative model for PHH.…”

Section: Introductionmentioning

confidence: 99%

“…HepaRG cells are bipotent progenitor cells and can differentiate into either biliary or hepatocyte lineages, representing the only example of complete differentiation in vitro (Cerec et al, 2007). As shown by genome-wide gene expression profiling studies, HepaRG cells are more similar to PHH and human liver tissue than any other liver cell line, particularly concerning the drug-processing genes (Hart et al, 2010;Rogue et al, 2012). In particular, HepaRG cells demonstrate stable expression of key phase 1 (e.g., P450s) and phase 2 (e.g., uridine diphosphate glucuronosyltransferases (UGTs) and glutathione S-transferases (GSTs)) enzymes, drug transporters (e.g., ATPbinding cassette transporters (ABCs) and solute carriers (SLCs)), and nuclear receptors (e.g., constitutive androstane receptor (CAR), pregnane X receptor (PXR), and peroxisome proliferator-activated receptors (PPARs)) (Aninat et al, 2006;Andersson et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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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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supporting

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…In addition, large interindividual variability has been observed in both the basal expression (Rogue et al, 2012) and in response to treatment of many genes related to drug metabolism. Liguori et al (2005) showed that, after 24 hours of treatment with trovafloxacin, gene changes varied across four different primary human hepatocyte (PHH) cultures.…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic Hepatotoxicity Signature of Chlorpromazine after Short- and Long-Term Exposure in Primary Human Sandwich Cultures

et al. 2013

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Drug-induced liver injury is the most frequent reason for market withdrawal of approved drugs, and is difficult to predict in animal models. Here, we analyzed transcriptomic data derived from shortand long-term cultured primary human hepatocytes (PHH) exposed to the well known human hepatotoxin chlorpromazine (CPZ). Samples were collected from five PHH cultures after short-term (1 and 3 days) and long-term (14 days) repeat daily treatment with 0.1 or 0.2 mM CPZ, corresponding to C max . Two PHH cultures were additionally treated with 1 mM CPZ, and the three others with 0.02 mM CPZ. Differences in the total number of gene changes were seen between donors and throughout treatment. Specific transcriptomic hepatotoxicity signatures were created for CPZ and consisted of inflammation/hepatitis, cholestasis, and liver proliferation in all five donors, as well as fibrosis and steatosis, which were observed in four of five donors. Necrosis was present in three of five donors, and an indicative signature of cirrhosis was observed after long-term 14-day repeat treatment, also in three of five donors. The inter-donor variability in the inflammatory response to CPZ treatment was associated with variability in the strength of the response of the transcriptomic hepatotoxicity signatures, suggesting that features of inflammation could be related to the idiosyncratic hepatotoxic effects of CPZ in humans.

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“…However, the use of fresh human hepatocytes has several drawbacks, such as scarce and unpredictable availability, interdonor variability, and significant variation in cell functions, especially cytochrome P450 (P450) activities (Luo et al, 2002;Ohtsuki et al, 2012;Rogue et al, 2012;Schaefer et al, 2012). Today, pooled cryopreserved human hepatocytes from several donors are used routinely in drug discovery and development to overcome the inconsistencies associated with fresh human hepatocytes.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Evaluation of Major In Vivo Drug Metabolic Pathways Using Primary Human Hepatocytes and HepaRG Cells in Suspension and a Dynamic Three-Dimensional Bioreactor System

Darnell

Ulvestad

Ellis

et al. 2012

J Pharmacol Exp Ther

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Major human specific metabolites, not detected during in vivo and in vitro preclinical studies, may cause unexpected drug interactions and toxicity in human and delays in clinical programs. Thus, reliable preclinical tools for the detection of major human metabolites are of high importance. The aim of this study was to compare major drug metabolic pathways in HepaRG cells, a human hepatoma cell line, to fresh human hepatocytes, cryopreserved human hepatocytes, and human in vivo data. Furthermore, the maintenance of cytochrome P450 (P450) and UDP-glucuronosyltransferase (UGT) activities in a dynamic three-dimensional (3D) bioreactor were evaluated over time by using HepaRG cells and human hepatocytes.14 Cdiclofenac and a candidate from AstraZeneca's drug development program, 14 C-AZD6610, which are metabolized by P450 and UGT in vivo, were used as model substrates. The proportion of relevant biotransformation pathways of the investigated drug was clearly different in the various cell systems. The hydroxylation route was favored in primary human hepatocytes, whereas the glucuronidation route was favored in HepaRG cells. The human in vivo metabolite profile of AZD6610 was best represented by human hepatocytes, whereas all major diclofenac metabolites were detected in HepaRG cells. Moreover, the metabolite profiles in cryopreserved and fresh human hepatocytes were essentially the same. The liver bioreactor using both fresh human hepatocytes and HepaRG cells retained biotransformation capacity over 1 week. Thus, the incubation time can be increased from a few hours in suspension to several days in 3D cultures, which opens up for detection of metabolites from slowly metabolized drugs.

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Interindividual Variability in Gene Expression Profiles in Human Hepatocytes and Comparison with HepaRG Cells

Cited by 58 publications

References 30 publications

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

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

Transcriptomic Hepatotoxicity Signature of Chlorpromazine after Short- and Long-Term Exposure in Primary Human Sandwich Cultures

In Vitro Evaluation of Major In Vivo Drug Metabolic Pathways Using Primary Human Hepatocytes and HepaRG Cells in Suspension and a Dynamic Three-Dimensional Bioreactor System

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