Optimization of the HepaRG cell model for drug metabolism and toxicity studies

Anthérieu, Sébastien; Chesné, Christophe; Li, Ruoya; Guguen‐Guillouzo, Christiane; Guillouzo, André

doi:10.1016/j.tiv.2012.05.008

Cited by 142 publications

(98 citation statements)

References 56 publications

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“…The metabolically competent human HepaRG cell-line represents a pertinent surrogate for primary human hepatocytes to investigate drug toxicity in vitro (Guillouzo 1998;Antherieu et al 2012). This cell-line has the capability to differentiate into two types of cells: hepatocyte-like colonies surrounded by clear primitive biliary cells.…”

Section: Introductionmentioning

confidence: 99%

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

et al. 2017

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Magnetic mesoporous silica nanoparticles (M-MSNs) are a promising class of nanoparticles for drug delivery. However, a deep understanding of the toxicological mechanisms of action of these nanocarriers is essential, especially in the liver. The potential toxicity on HepaRG cells of pristine, pegylated (PEG), and lipid (DMPC) M-MSNs were compared. Based on MTT assay and real-time cell impedance, none of these NPs presented an extensive toxicity on hepatic cells. However, we observed by transmission electron microscopy (TEM) that the DMPC and pristine M-MSNs were greatly internalized. In comparison, PEG M-MSNs showed a slower cellular uptake. Whole gene expression profiling revealed the M-MSNs molecular modes of action in a time-and dose-dependent manner. The lowest dose tested (1.6 mg/cm 2 ) induced no molecular effect and was defined as 'No Observed Transcriptional Effect level.' The dose 16 mg/cm 2 revealed nascent but transient effects. At the highest dose (80 mg/cm 2 ), adverse effects have clearly arisen and increased over time. The limit of biocompatibility for HepaRG cells could be set at 16 mg/cm 2 for these NPs. Thanks to a comparative pathway-driven analysis, we highlighted the sequence of events that leads to the disruption of hepatobiliary system, elicited by the three types of MMSNs, at the highest dose. The Adverse Outcome Pathway of hepatic cholestasis was implicated. Toxicogenomics applied to cell cultures is an effective tool to characterize and compare the modes of action of many substances. We propose this strategy as an asset for upstream selection of the safest nanocarriers in the framework of regulation for nanobiosafety. ARTICLE HISTORY

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Section: Introductionmentioning

confidence: 99%

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

et al. 2017

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“…Global profiles of transporter regulation by PMA in HepaRG cells and human hepatocytes were however significantly correlated, as well as those of PKC isoform 18 expression. In the same way, various similarities in detoxifying pathway expressions and regulations between HepaRG cells and human hepatocytes have been reported [54]. The overall and now-well established interest of HepaRG cells as a convenient alternative to the use of human hepatocytes in pharmacological and/or toxicological areas [55] has therefore not to be challenged, even if some differences between these two in vitro models have likely to be kept in mind.…”

Section: Discussionmentioning

confidence: 88%

Protein kinase C-dependent regulation of human hepatic drug transporter expression

Mayati

Vée

Moreau

et al. 2015

Biochemical Pharmacology

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Hepatic drug transporters are now recognized as major actors of hepatobiliary elimination of drugs. Characterization of their regulatory pathways is therefore an important issue. In this context, the present study was designed to analyze the potential regulation of human hepatic transporter expression by protein kinase C (PKC) activation. Treatment by the reference PKC activator phorbol 12-myristate 13-acetate (PMA) for 48h was shown to decrease mRNA expression of various sinusoidal transporters, including OATP1B1, OATP2B1, NTCP, OCT1 and MRP3, but to increase that of OATP1B3, whereas mRNA expression of canalicular transporters was transiently enhanced (MDR1), decreased (BSEP and MRP2) or unchanged (BCRP) in human hepatoma HepaRG cells. The profile of hepatic transporter mRNA expression changes in PMA-treated HepaRG cells was correlated to that found in PMA-exposed primary human hepatocytes and was similarly observed in response to the PKC-activating marketed drug ingenol mebutate. It was associated with concomitant repression of OATP1B1 and OATP2B1 protein expression and reduction of OATP, OCT1, NTCP and MRP2 activity. The use of chemical PKC inhibitors further suggested a contribution of novel PKCs isoforms to PMA-mediated regulations of transporter mRNA expression. PMA was finally shown to cause epithelial-mesenchymal transition (EMT) in HepaRG cells and exposure to various additional EMT inducers, i.e., hepatocyte growth factor, tumor growth factor-1 or the HNF4 inhibitor BI6015, led to transporter expression alterations highly correlated to those triggered by PMA. Taken together, these data highlight PKC-dependent regulation of human hepatic drug transporter expression, which may be closely linked to EMT triggered by PKC activation.

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“…1B). DMSO-treated differentiated HepaRG cells are well-known to be polarized (Antherieu et al, 2012) and consequently exhibit bile canaliculi-like structures, which are fully functional because they accumulated the fluorescent dye CF (Fig. 2A).…”

Section: Resultsmentioning

confidence: 99%

“…Indeed, when cultured in appropriated conditions, i.e., in the presence of 2 % (vol/vol) dimethylsulfoxide (DMSO), HepaRG cells express high levels of hepatic drug detoxifying pathways, including phase 1 enzymes, such as cytochromes P-450 1A2, 2B6 and 3A4, and phase 2 enzymes such as such glutathione S-transferases A1/A2, A4 and M1 and UDPglucuronosyl transferase 1A1 (Aninat et al, 2006;Antherieu et al, 2010). Moreover, HepaRG cells exhibit notable expression of drug-sensing receptors such as pregnane X receptor and constitutive androstane receptor (Antherieu et al, 2012) and are consequently responsive to inducers of drug metabolism (Kanebratt and Andersson, 2008;Turpeinen et al, 2009). …”

Section: Introductionmentioning

confidence: 99%

Differential regulation of drug transporter expression by all-trans retinoic acid in hepatoma HepaRG cells and human hepatocytes

Vée

Jouan

Stieger³

et al. 2013

European Journal of Pharmaceutical Sciences

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All-trans retinoic acid (atRA) is the active form of vitamin A, known to activate retinoid receptors, especially the heterodimer retinoid X receptor (RXR):retinoic acid receptor (RAR) that otherwise may play a role in regulation of some drug transporters. The present study was designed to characterize the nature of human hepatic transporters that may be targeted by atRA and the heterodimer RXR:RAR. Exposure of human hepatoma HepaRG cells and primary human hepatocytes to 5 M atRA down-regulated mRNA levels of various sinusoidal solute carrier (SLC) influx transporters, including organic anion transporting polypeptide (OATP) 2B1, OATP1B1, organic cation transporter (OCT) 1 and organic anion transporter (OAT) 2, and induced those of the canalicular breast cancer resistance protein (BCRP). The retinoid concomitantly reduced protein expression of OATP2B1 and OATP1B1 and activity of OATPs and OCT1 and induced BCRP protein expression in HepaRG cells. Some transporters such as OATP1B3 and the bile salt export pump (BSEP) were however down-regulated by atRA in primary human hepatocytes, but induced in HepaRG cells, thus pointing out discrepancies between these two liver cell models in terms of detoxifying protein regulation. atRA-mediated repressions of OATP2B1, OATP1B1, OAT2 and OCT1 mRNA expression were finally shown to be counteracted by knocking-down expression of RAR and RXR through siRNA transfection in HepaRG cells. atRA thus differentially regulated human hepatic drug transporters, mainly in a RXR:RAR-dependent manner, therefore establishing retinoids and retinoid receptors as modulators of liver drug transporter expression.

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Optimization of the HepaRG cell model for drug metabolism and toxicity studies

Cited by 142 publications

References 56 publications

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

Protein kinase C-dependent regulation of human hepatic drug transporter expression

Differential regulation of drug transporter expression by all-trans retinoic acid in hepatoma HepaRG cells and human hepatocytes

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