title: Human embryonic stem cell derived hepatocyte-like cells as a tool for in vitro hazard assessment of chemical carcinogenicity.
abstract: Hepatocyte-like cells derived from the differentiation of human embryonic stem cells (hES-Hep) have potential to provide a human relevant in vitro test system in which to evaluate the carcinogenic hazard of chemicals. In this study, we have investigated this potential using a panel of 15 chemicals classified as noncarcinogens, genotoxic carcinogens, and nongenotoxic carcinogens and measured whole-genome transcriptome responses with gene expression microarrays. We applied an ANOVA model that identified 592 genes highly discriminative for the panel of chemicals. Supervised classification with these genes achieved a cross-validation accuracy of > 95%. Moreover, the expression of the response genes in hES-Hep was strongly correlated with that in human primary hepatocytes cultured in vitro. In order to infer mechanistic information on the consequences of chemical exposure in hES-Hep, we developed a computational method that measures the responses of biochemical pathways to the panel of treatments and showed that these responses were discriminative for the three toxicity classes and linked to carcinogenesis through p53, mitogen-activated protein kinases, and apoptosis pathway modules. It could further be shown that the discrimination of toxicity classes was improved when analyzing the microarray data at the pathway level. In summary, our results demonstrate, for the first time, the potential of human embryonic stem cell--derived hepatic cells as an in vitro model for hazard assessment of chemical carcinogenesis, although it should be noted that more compounds are needed to test the robustness of the assay.
authors.affiliation: Department of Vertebrate Genomics, Max Planck Institute for Molecular Genetics, D-14195 Berlin, Germany.
authors.forename: Reha
authors.initials: R
authors.lastname: Yildirimman
authors.forename: Gabriella
authors.initials: G
authors.lastname: Brolén
authors.forename: Mireia
authors.initials: M
authors.lastname: Vilardell
authors.forename: Gustav
authors.initials: G
authors.lastname: Eriksson
authors.forename: Jane
authors.initials: J
authors.lastname: Synnergren
authors.forename: Hans
authors.initials: H
authors.lastname: Gmuender
authors.forename: Atanas
authors.initials: A
authors.lastname: Kamburov
authors.forename: Magnus
authors.initials: M
authors.lastname: Ingelman-Sundberg
authors.forename: José
authors.initials: J
authors.lastname: Castell
authors.forename: Agustin
authors.initials: A
authors.lastname: Lahoz
authors.forename: Jos
authors.initials: J
authors.lastname: Kleinjans
authors.forename: Joost
authors.initials: J
authors.lastname: van Delft
authors.forename: Petter
authors.initials: P
authors.lastname: Björquist
authors.forename: Ralf
authors.initials: R
authors.lastname: Herwig
mesh_terms: D000704:Analysis of Variance; D015197:Carcinogenicity Tests / Q000379:methods*; D002273:Carcinogens / Q000633:toxicity*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019295:Computational Biology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D015386:Hazardous Substances / Q000633:toxicity*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D046228:Microarray Analysis; D008858:Microscopy, Phase-Contrast; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D002273:Carcinogens; D015386:Hazardous Substances; D003577:Cytochrome P-450 Enzyme System