title: Validation of precision-cut liver slices to study drug-induced cholestasis: a transcriptomics approach.
abstract: Hepatotoxicity is one of the major reasons for withdrawal of drugs from the market. Therefore, there is a need to screen new drugs for hepatotoxicity in humans at an earlier stage. The aim of this study was to validate human precision-cut liver slices (PCLS) as an ex vivo model to predict drug-induced cholestasis and identify the possible mechanisms of cholestasis-induced toxicity using gene expression profiles. Five hepatotoxicants, which are known to induce cholestasis (alpha-naphthyl isothiocyanate, chlorpromazine, cyclosporine, ethinyl estradiol and methyl testosterone) were used at concentrations inducing low (<30 %) and medium (30-50 %) toxicity, based on ATP content. Human PCLS were incubated with the drugs in the presence of a non-toxic concentration (60 µM) of a bile acid mixture (portal vein concentration and composition) as model for bile acid-induced cholestasis. Regulated genes include bile acid transporters and cholesterol transporters. Pathway analysis revealed that hepatic cholestasis was among the top ten regulated pathways, and signaling pathways such as farnesoid X receptor- and liver X receptor-mediated responses, which are known to play a role in cholestasis, were significantly affected by all cholestatic compounds. Other significantly affected pathways include unfolded protein response and protein ubiquitination implicating the role of endoplasmic reticulum stress. This study shows that human PCLS incubated in the presence of a physiological bile acid mixture correctly reflect the pathways affected in drug-induced cholestasis in the human liver. In the future, this human PCLS model can be used to identify cholestatic adverse drug reactions of new chemical entities.
authors.affiliation: Division of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands.
authors.forename: Suresh
authors.initials: S
authors.lastname: Vatakuti
authors.affiliation: Division of Pharmaceutical Technology and Biopharmacy, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Olinga
authors.affiliation: National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Jeroen L A
authors.initials: JLA
authors.lastname: Pennings
authors.affiliation: Division of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands. g.m.m.groothuis@rug.nl.
authors.forename: Geny M M
authors.initials: GMM
authors.lastname: Groothuis
mesh_terms: D015058:1-Naphthylisothiocyanate / Q000633:toxicity; D000368:Aged; D002746:Chlorpromazine / Q000009:adverse effects; D002779:Cholestasis / Q000139:chemically induced* / Q000235:genetics; D016572:Cyclosporine / Q000009:adverse effects; D004305:Dose-Response Relationship, Drug; D004997:Ethinyl Estradiol / Q000009:adverse effects; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D008099:Liver / Q000187:drug effects*; D008297:Male; D008777:Methyltestosterone / Q000009:adverse effects; D008875:Middle Aged; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D059467:Transcriptome / Q000187:drug effects; D055815:Young Adult
chemical_list: D004997:Ethinyl Estradiol; D015058:1-Naphthylisothiocyanate; D016572:Cyclosporine; D002746:Chlorpromazine; D008777:Methyltestosterone
keywords: Cholestasis; Hepatotoxicity; Precision-cut liver slices; Transcriptomics