Establishment of a Drug-Induced, Bile Acid–Dependent Hepatotoxicity Model Using HepaRG Cells

Susukida, Takeshi; Sekine, Satoshi; Nozaki, Mayuka; Tokizono, Mayuko; Oizumi, Kumiko; Horie, Toshiharu; Ito, Kousei

doi:10.1016/j.xphs.2016.01.013

Cited by 26 publications

(17 citation statements)

References 49 publications

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“…Whilst valuable, there is still a need for further mechanistic insight into the pathology of DIC that would require an in vitro model with enhanced physiology. HepaRG cells have been postulated to be an improved model choice for DIC studies (Hendriks et al, 2016; Susukida et al, 2016; Woolbright et al, 2016). HepaRG cells are a bipotent progenitor cell line, capable of differentiating into hepatocytes and primitive biliary-like cells.…”

Section: Introductionmentioning

confidence: 99%

Differential toxic effects of bile acid mixtures in isolated mitochondria and physiologically relevant HepaRG cells

Penman

Sharma

Aerts

et al. 2019

Toxicology in Vitro

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Bile acids (BAs) are recognised as the causative agents of toxicity in drug-induced cholestasis (DIC). Research in isolated mitochondria and HepG2 cells have demonstrated BA-mediated mitochondrial dysfunction as a key mechanism of toxicity in DIC. However, HepG2 cells are of limited suitability for DIC studies as they do not express the necessary physiological characteristics. In this study, the mitotoxic potentials of BA mixtures were assessed in isolated mitochondria and a better-suited hepatic model, HepaRG cells. BAs induced structural alterations and a loss of mitochondrial membrane potential (MMP) in isolated mitochondria however, this toxicity did not translate to HepaRG cells. There were no changes in oxygen consumption rate, MMP or ATP levels in glucose and galactose media, indicating that there was no direct mitochondrial toxicity mediated via electron transport chain dysfunction in HepaRG cells. Assessment of key biliary transporters revealed that there was a time-dependent reduction in the expression and activity of multi-drug resistance protein 2 (MRP2), which was consistent with the induction of cytotoxicity in HepaRG cells. Overall, the findings from this study have demonstrated that mitochondrial dysfunction is not a mechanism of BA-induced toxicity in HepaRG cells.

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Section: Introductionmentioning

confidence: 99%

Differential toxic effects of bile acid mixtures in isolated mitochondria and physiologically relevant HepaRG cells

Penman

Sharma

Aerts

et al. 2019

Toxicology in Vitro

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“…We previously reported a unique cell-based toxicity assay using sandwich-cultured hepatocytes (SCH) in combination with a titrated amount of human BA species. 10 If a test drug inhibits BA efflux from SCH, the accumulation of BAs eventually induces cell death, 11,12 and BA-dependent hepatocyte toxicity (BA tox ) is detected by the release of lactate dehydrogenase (LDH). The in vitro assay system successfully separated DILI high-risk and low-risk drugs in the clinical situation with more than about 80% sensitivity and specificity.…”

Section: Introductionmentioning

confidence: 99%

Metabolic Activation of Cholestatic Drug-Induced Bile Acid-Dependent Toxicity in Human Sandwich-Cultured Hepatocytes

Ogimura

Tokizono

Sekine

et al. 2017

Journal of Pharmaceutical Sciences

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“…HepaRG cells express ZIP14 and the transporter localizes to the basolateral surface HepaRG cells polarize to mature hepatocytes and form confluent monolayers with bile canalicular formations. They have been utilized to study various transport mechanisms and have successfully been imaged using indirect immunofluorescence (Antherieu et al 2013;Bachour-El Azzi et al 2015;Kanebratt and Andersson 2008;Sharanek et al 2014;Susukida et al 2016). To determine the expression and membrane distribution of ZIP14, we performed indirect immunofluorescence experiments co-labeling apical or basolateral markers.…”

Section: Resultsmentioning

confidence: 99%

ZIP14 is degraded in response to manganese exposure

Thompson

Wessling‐Resnick

2019

Biometals

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Manganese (Mn) is an essential element necessary for proper development and brain function. Circulating Mn levels are regulated by hepatobiliary clearance to limit toxic levels and prevent tissue deposition. To characterize mechanisms involved in hepatocyte Mn uptake, polarized human HepaRG cells were used for this study. Western blot analysis and immunofluorescence microscopy showed the Mn transporter ZIP14 was expressed and localized to the basolateral surface of polarized HepaRG cells. HepaRG cells took up 54Mn in a time- and temperature-dependent manner but uptake was reduced after exposure to Mn. This loss in transport activity was associated with decreased ZIP14 protein levels in response to Mn exposure. Mn-induced degradation of ZIP14 was blocked by bafilomycin A1, which increased localization of the transporter in Lamp1-positive vesicles. Mn exposure also down-regulated the Golgi proteins TMEM165 and GPP130 while the ER stress marker BiP was induced. These results indicate that Mn exposure decreases ZIP14 protein levels to limit subsequent uptake of Mn as a cytoprotective response. Thus, high levels of Mn may compromise first-pass-hepatic clearance mechanisms.

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Establishment of a Drug-Induced, Bile Acid–Dependent Hepatotoxicity Model Using HepaRG Cells

Cited by 26 publications

References 49 publications

Differential toxic effects of bile acid mixtures in isolated mitochondria and physiologically relevant HepaRG cells

Differential toxic effects of bile acid mixtures in isolated mitochondria and physiologically relevant HepaRG cells

Metabolic Activation of Cholestatic Drug-Induced Bile Acid-Dependent Toxicity in Human Sandwich-Cultured Hepatocytes

ZIP14 is degraded in response to manganese exposure

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