Utility of B-13 Progenitor-Derived Hepatocytes in Hepatotoxicity and Genotoxicity Studies

Probert, Philip M. E.; Chung, Git; Cockell, Simon; Agius, Loranne; Mosesso, Pasquale; White, Steven A.; Oakley, Fiona; Brown, Charles D.; Wright, Matthew C.

doi:10.1093/toxsci/kft258

Cited by 18 publications

(71 citation statements)

References 52 publications

(71 reference statements)

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“…5 B and C demonstrate that methapyrilene exposure resulted in phospholipidosis in both B-13 and B-13/H cells in dose-dependent manners, most potently in B-13 cells. As previously reported ( Probert et al, 2014 ), methapyrilene was only toxic to B-13/H cells and not to B-13 cells and similar results were observed in these studies ( Fig. 5 D).…”

Section: Resultssupporting

confidence: 92%

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B-13 progenitor-derived hepatocytes (B-13/H cells) model lipid dysregulation in response to drugs and chemicals

et al. 2017

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Lipid dysregulation is a common hepatic adverse outcome after exposure to toxic drugs and chemicals. A donor-free rat hepatocyte-like (B-13/H) cell was therefore examined as an in vitro model for investigating mechanisms. The B-13/H cell irreversibly accumulated triglycerides (steatosis) in a time- and dose-dependent manner when exposed to fatty acids, an effect that was potentiated by the combined addition of hyperglycaemic levels of glucose and insulin. B-13/H cells also expressed the LXR nuclear receptors and exposure to their activators – T0901317 or GW3965 – induced luciferase expression from a transfected LXR-regulated reporter gene construct and steatosis in a dose-dependent manner with T0901317. Exposing B-13/H cells to a variety of cationic amphiphilic drugs – but not other hepatotoxins – also resulted in a time- and dose-dependent accumulation of phospholipids (phospholipidosis), an effect that was reduced by over-expression of lysosomal phospholipase A2. Through application of this model, hepatotoxin methapyrilene exposure was shown to induce phospholipidosis in both B-13 and B-13/H cells in a time- and dose-dependent manner. However, methapyrilene was only toxic to B-13/H cells and inhibitors of hepatotoxicity enhanced phospholipidosis, suggesting phospholipidosis is not a pathway in toxicity for this withdrawn drug. In contrast, pre-existing steatosis had minimal effect on methapyrilene hepatotoxicity in B-13/H cells. These data demonstrate that the donor free B-13 cell system for generating hepatocyte-like cells may be employed in studies of fatty acid- and LXR activator-induced steatosis and phospholipidosis and in the dissection of pathways leading to adverse outcomes such as hepatotoxicity.

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Section: Resultssupporting

confidence: 92%

“…5 D). Previous work has also demonstrated that methapyrilene hepatoxicity is inhibited by sulfhydryl reducing agents and the Ca 2+ channel blocker nifedipine ( Ratra et al, 1998a , Probert et al, 2014 ). Fig.…”

Section: Resultsmentioning

confidence: 97%

B-13 progenitor-derived hepatocytes (B-13/H cells) model lipid dysregulation in response to drugs and chemicals

et al. 2017

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“…The transdifferentiated rat B13/H cell system has been reported to express a range of constitutive and inducible drug metabolising enzymes, such as the Cyp enzymes (Probert et al 2014). Therefore the hepatocyte-like B13/H cells were subsequently treated with PB (2mM) for up to 9 days, with the treatment and media replenished every 72 h. Analysis of the treated cells demonstrated significant increases in Cyp2b1 mRNA ( Figure 2A) and Cyp2b activity across the 9 days ( Figure 2B).…”

Section: Phenobarbital Treatmentmentioning

confidence: 99%

“…It was reported by Shen et al, (Shen et al 2000) that treatment of the undifferentiated B13 cell with dexamethasone for two weeks stimulated the cell to transdifferentiate into hepatocyte-like B13/H cells. The differentiated cell population display gene expression profiles similar to primary hepatocytes and can be maintained in culture for considerably longer than primary hepatocytes without de-differentiation (Probert et al 2013;Probert et al 2015), making them a useful model for the proposed treatments here.…”

Section: Introductionmentioning

confidence: 99%

Transdifferentiated rat pancreatic progenitor cells (AR42J-B13/H) respond to phenobarbital in a rat hepatocyte-specific manner.

et al. 2016

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Phenobarbital (PB) is known to produce species-specific effects in the rat and mouse, being carcinogenic in certain mouse strains, but only in rats if treated after a DNA damaging event.PB treatment in the rat and mouse also produces disparate effects on cell signalling and miRNA expression profiles. These responses are induced by short term and prolonged PB exposure, respectively, with the latter treatments being difficult to examine mechanistically in primary hepatocytes due to rapid loss of the original hepatic phenotype and limited sustainability in culture. Here we explore the rat hepatocyte-like B13/H cell line as a model for hepatic response to PB exposure in both short-term and longer duration treatments. We demonstrate that PB with Egf treatment in the B13/H cells, resulted in a significant increase in Erk activation, as determined by the ratio of phospho-Erk to total Erk, compared to Egf alone. We also show that an extended treatment with PB in the B13/H cells produces a miRNA response similar to that seen in the rat in vivo, via the time-dependent induction of miR-182/96. Additionally, we confirm that B13/H cells respond to Car activators in a typical rat-specific manner. These data suggest that the B13/H cells produce temporal responses to PB that are comparable to those reported in short-term primary rat hepatocyte cultures and in the longer term are similar to those in the rat in vivo. Finally, we also show that Carassociated miR-122 expression is decreased by PB treatment in B13/H cells, a PB-induced response that is common to the rat, mouse and human. We conclude that the B13/H cell system produces a qualitative response comparable to the rat, which is different to the response in the mouse, and that this model could be a useful tool for exploring the functional consequences of PB-sensitive miRNA changes and resistance to PB-mediated tumours in the rat.

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“…On the other hand, qPCRbased CARCINOscreen ® evaluation can be made by comparing the expression ratios of any of the four predictive genes with the threshold values. With regard to the four selected predictive genes for qPCR, Abcb1b is a member of the ATP-binding cassette (ABC) protein superfamily and multi-drug resistanceassociated protein, and the protein transporter is reported as being functional in B-13/H cancer cells (Probert et al, 2014). Merrick et al (2012) reported that administration of the hepatocarcinogen aflatoxin B1 (AFB1) to male F344 rats significantly increased the expression of Abcb1b in the rat liver.…”

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confidence: 99%

Simpler alternative to CARCINOscreen<sup>®</sup> based on quantitative PCR (qPCR)

et al. 2016

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-Carcinogenicity of chemicals in our environment is one of the most important health hazards to humans. Recently, a microarray-based short-term prediction system for the hepatocarcinogenicity of chemicals, named CARCINOscreen ® , was developed. Although the system is a promising tool reported to have an ability to predict hepatocarcinogenicity in rats with 92.9% accuracy, it requires specialized equipment and skilled bioinformatics approaches for data analysis. Therefore, we attempted to develop a quantitative PCR (qPCR)-based system as an alternative to microarray-based CARCINOscreen ® . Finally, an optimized gene set consisting of four predictive genes (Abcb1b, Eprs, Map3K8, and Igh-6) was selected from among 3,150 combinations of candidate gene sets. The results of training-and validation-phase trials showed that the qPCR-based alternative to the microarray-based CARCINOscreen ® could predict the hepatocarcinogenicity of chemicals in rats with 82.8%-86.4% accuracy. One of the predictive genes, Abcb1b, a member of the ATP-binding cassette protein superfamily and multi-drug resistance-associated protein, and the results of this study may indicate a close relation of this gene to the carcinogenicity of chemicals. The prediction performance of the qPCR-based CARCINOscreen ® , as well as its user-friendliness and cost efficiency, suggests that this method is promising for application to primary health hazard assessment. Thus, the qPCR-based CARCINOscreen ® is considered as a promising tool for predicting the carcinogenicity of chemicals.

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Utility of B-13 Progenitor-Derived Hepatocytes in Hepatotoxicity and Genotoxicity Studies

Cited by 18 publications

References 52 publications

B-13 progenitor-derived hepatocytes (B-13/H cells) model lipid dysregulation in response to drugs and chemicals

B-13 progenitor-derived hepatocytes (B-13/H cells) model lipid dysregulation in response to drugs and chemicals

Transdifferentiated rat pancreatic progenitor cells (AR42J-B13/H) respond to phenobarbital in a rat hepatocyte-specific manner.

Simpler alternative to CARCINOscreen<sup>®</sup> based on quantitative PCR (qPCR)

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