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

Lookup NU author(s): Dr Phillip Probert, Git Chung, Dr Simon CockellORCiD, Emerita Professor Loranne Agius, Steven White, Professor Fiona OakleyORCiD, Dr Colin Brown, Professor Matthew Wright

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Abstract

AR42J B-13 (B-13) cells form hepatocyte-like (B-13/H) cells in response to glucocorticoid treatment. To establish its utility in toxicity and genotoxicity screening, CYP induction; susceptibility to toxins and transporter gene expression were examined. Conversion to B-13/H cells resulted in expression of male-specific CYP2C11 and sensitivity to methapyrilene. B-13/H cells constitutively expressed CYP1A, induced expression in response to an AhR agonist and activated benzo[α]yrene to a DNA damaging species. Functional CYP1A2 was not expressed due to deletions in the CYP1A2 gene. A B-13 cell line stably expressing the human CYP1A2 was therefore engineered (B-13^-TR/h1A2) and thederived B-13/H cells expressed metabolically functional CYP1A2. Treatment with the cooked food mutagen 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine resulted in a dose-dependent increase in DNA damage. B-13/H cells expressed CAR and induced CYP2B1 mRNA levels in response to classical CAR activators. However, translation to functional CYP2B1 protein was low and increased minimally by CAR activator treatment. B-13/H cells expressed high levels of PXR and induced CYP3A1 in response to classical PXR activators. CYP3A genes were inducible, functional and activated aflatoxin B1 to a DNA damaging species. All 23 major hepatic transporters were induced when B-13 cells were converted to B-13/H cells, although in many cases, levels remained below those present in adult rat liver. However, BSEP, Abcb1b, MRP and BCRP transporters were functional in B-13/H cells. These data demonstrate that the B-13 cell generates hepatocyte-like cells with functional drug metabolism and transporter activities which can alone - or in a humanised form - be used to screen for hepatotoxic and genotoxic endpoints in vitro.

Publication metadata

Author(s): Probert PM, Chung GW, Cockell SJ, Agius L, Mosesso P, White SA, Oakley F, Brown CD, Wright MC

Publication type: Article

Publication status: Published

Journal: Toxicological Sciences

Year: 2014

Volume: 137

Issue: 2

Pages: 350-370

Print publication date: 01/02/2014

Online publication date: 14/11/2013

Acceptance date: 04/11/2013

Date deposited: 11/02/2014

ISSN (print): 1096-6080

ISSN (electronic): 1096-0929

Publisher: Oxford University Press

URL: http://dx.doi.org/10.1093/toxsci/kft258

DOI: 10.1093/toxsci/kft258

PubMed id: 24235770

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