title: Cyclosporine A treated in vitro models induce cholestasis response through comparison of phenotype-directed gene expression analysis of in vivo Cyclosporine A-induced cholestasis.
abstract: In vitro models for hepatotoxicity testing are a necessity for advancement of toxicological research. Assessing the in vitro response requires in vivo validated gene sets reflective of the hepatotoxic phenotype. Cholestasis, the impairment of bile flow, is induced in C57BL/6J mice treated with cyclosporine A (CsA) to identify phenotype reflective gene sets. CsA treatment through oral gavage for 25 days induced cholestasis, as confirmed by histopathology and serum chemistry. Over 1, 4, and 11 days of CsA exposure gradual increases in serum markers were correlated to gene expression. This phenotype-directed analysis identified gene sets specific to the onset and progression of cholestasis, such as PPAR related processes and drug metabolism, by circumventing other effects of CsA, such as immunosuppression, found in dose*time group analysis. In vivo gene sets are enriched in publicly available data sets of CsA-treated HepaRG and primary mouse hepatocytes. However, genes identified within these gene sets did not overlap between in vivo and in vitro. In vitro regulated genes represent the initial response to cholestasis, whereas in vivo genes represent the later adaptive response. We conclude that the applicability of in vitro models for hepatotoxicity testing fully depends on a solid in vivo phenotype anchored analysis.
authors.affiliation: Center for Health Protection, National Institute for Public Health and the Environment-RIVM, 3720 BA, Bilthoven, The Netherlands.
authors.forename: Anne S
authors.initials: AS
authors.lastname: Kienhuis
authors.forename: Alexa P
authors.initials: AP
authors.lastname: Vitins
authors.forename: Jeroen L A
authors.initials: JL
authors.lastname: Pennings
authors.forename: Tessa E
authors.initials: TE
authors.lastname: Pronk
authors.forename: Ewoud N
authors.initials: EN
authors.lastname: Speksnijder
authors.forename: Marianne
authors.initials: M
authors.lastname: Roodbergen
authors.forename: Joost H M
authors.initials: JH
authors.lastname: van Delft
authors.forename: Mirjam
authors.initials: M
authors.lastname: Luijten
authors.forename: Leo T M
authors.initials: LT
authors.lastname: van der Ven
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D001647:Bile Acids and Salts / Q000097:blood; D001663:Bilirubin / Q000097:blood; D002779:Cholestasis / Q000097:blood / Q000139:chemically induced* / Q000235:genetics; D002784:Cholesterol / Q000097:blood; D016572:Cyclosporine / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling / Q000379:methods; D056945:Hep G2 Cells; D006651:Histocytochemistry; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010641:Phenotype; D013047:Specific Pathogen-Free Organisms; D043922:Toxicogenetics
chemical_list: D001647:Bile Acids and Salts; D016572:Cyclosporine; D002784:Cholesterol; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D001663:Bilirubin
keywords: Cholestasis; Cyclosporin A; Hepatotoxicity; In vitro models; Mouse in vivo; Phenotype-directed gene expression analysis