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Classification of cholestatic and necrotic hepatotoxicants using transcriptomics on human precision-cut liver slices Suresh Vatakuti, Jeroen L. A. Pennings, Emilia Gore, Peter Olinga, and Geny M.M. Groothuis Chem. Res. Toxicol., Just Accepted Manuscript • DOI: 10.1021/acs.chemrestox.5b00491 • Publication Date (Web): 16 Feb 2016 Downloaded from http://pubs.acs.org on February 17, 2016
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Chemical Research in Toxicology
Classification of cholestatic and necrotic hepatotoxicants using transcriptomics on human precision-cut liver slices Suresh Vatakuti1, Jeroen L.A. Pennings3, Emilia Gore2, Peter Olinga2, Geny M. M. Groothuis1* 1
Division of Pharmacokinetics, Toxicology and Targeting, 2Division of Pharmaceutical Technology and Biopharmacy, Groningen Research Institute for Pharmacy, University of Groningen, Groningen and 3National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
Keywords: Classification, Hepatotoxicity, precision cut liver slices, cholestasis, necrosis
Corresponding author: *
Division of Pharmacokinetics, Toxicology and Targeting,
Department of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. E-mail:
[email protected] ACS Paragon Plus Environment
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Chemical Research in Toxicology
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TOC Graphics
Preparation of human precision cut liver slices (3 PCLS/conc/liver) Exposure to cholestatic and necrotic compounds Gene expression data Predictive model development Support vector machine (SVM)
Random Forest (RF)
Leave-one-compound out validation Classifer genes identification
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Chemical Research in Toxicology
Abstract Human toxicity screening is an important stage in the development of safe drug candidates. Hepatotoxicity is one of the major reasons for withdrawal of drugs from the market because the liver is the major organ involved in drug metabolism and it can generate toxic metabolites. There is a need to screen molecules for drug-induced hepatotoxicity in humans at an earlier stage. Transcriptomics is a technique widely used to screen molecules for toxicity and to unravel toxicity mechanisms. To date the majority of such studies were performed using animals or animal cells, with concomitant difficulty in interpretation due to species differences, or in human hepatoma cell lines or cultured hepatocytes, suffering from the lack of physiological expression of enzymes and transporters and lack of non-parenchymal cells. The aim of this study was to classify known hepatotoxicants on their phenotype of toxicity in man using gene expression profiles ex vivo in human precision-cut liver slices (PCLS). Hepatotoxicants known to induce either necrosis (n=5) or cholestasis (n=5) were used at concentrations inducing low (
