Transcriptomics-based identification of developmental toxicants through their interference with cardiomyocyte differentiation of embryonic stem cells.

D.A. van Dartel, J.L. Pennings, F.J. van Schooten, A.H. Piersma

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    The embryonic stem cell test (EST) predicts developmental toxicity based on the inhibition of cardiomyocyte differentiation of embryonic stem cells (ESC). The subjective endpoint, the long culture duration together with the undefined applicability domain and related predictivity need further improvement to facilitate implementation of the EST into regulatory strategies. These aspects may be improved by studying gene expression changes in the ESC differentiation cultures and their modulation by compound exposure using transcriptomics. Here, we tested the developmental toxicants monobutyl phthalate and 6-aminonicotinamide. ESC were allowed to differentiated, and cardiomyocyte differentiation was assessed after 10 days of culture. RNA of solvent controls was collected after 0, 24, 48, 72 and 96 hours of exposure, and RNA of developmental-toxicant-exposed cultures was collected after 24 and 96 hours. Samples were hybridized to DNA microarrays, and 1355 genes were found differentially expressed among the unexposed experimental groups. These regulated genes were involved in differentiation-related processes, and Principal Component Analysis (PCA) based on these genes showed that the unexposed experimental groups appeared in chronological order in the PCA, which can therefore be regarded as a continuous representation of the differentiation track. The developmental-toxicant-exposed cultures appeared to deviate significantly from this differentiation track, which confirms the compound-modulating effects on the differentiation process. The incorporation of transcriptomics in the EST is expected to provide a more informative and improved endpoint in the EST as compared with morphology, allowing early detection of differentiation modulation. Furthermore, this approach may improve the definition of the applicability domain and predictivity of the EST.
    Original languageEnglish
    Pages (from-to)420-428
    Number of pages9
    JournalToxicology and Applied Pharmacology
    Volume243
    Issue number3
    DOIs
    Publication statusPublished - 15 Mar 2010

    Keywords

    • Embryonic stem cells
    • Alternative test method
    • Differentiation
    • Gene expression
    • Monobutyl phthalate
    • 6-Aminonicotinamide
    • N-BUTYL PHTHALATE
    • VITRO EMBRYOTOXICITY TESTS
    • GENE-EXPRESSION CHANGES
    • IN-VITRO
    • MOLECULAR SIGNATURE
    • DOWNSTREAM TARGET
    • ES CELLS
    • TOXICITY
    • RATS
    • PLURIPOTENCY

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