Monitoring developmental toxicity in the embryonic stem cell test using differential gene expression of differentiation-related genes.

D.A. van Dartel, J.L. Pennings, L.J. de la Fonteyne, M.H.M. van Herwijnen, J.H.M. van Delft, F.J. van Schooten, A.H. Piersma

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    The Embryonic Stem cell Test (EST) has been designed to predict developmental toxicity based upon compound-induced inhibition of embryonic stem cell (ESC) differentiation. The endpoint scoring, the test duration, and the definition of the predictivity and the applicability domain require improvements to facilitate implementation of the EST into regulatory testing strategies. The use of transcriptomics to study compound-induced differentiation modulation may improve the EST in each of these aspects. ESC differentiation was induced and cell samples were collected after 0, 24 and 48h of differentiation. Additionally, samples were collected that were 24h-exposed to one of five developmentally-toxic compounds or a non-developmentally-toxic compound. All samples were hybridized to Affymetrix GeneChips, and analyses revealed that 26 genes were significantly regulated both during ESC differentiation and by exposure to each of the developmentally-toxic compounds tested. Using Principal Component Analysis we defined a 'differentiation track' on the basis of this gene list, which represents ESC differentiation. We showed that significant deviation from the differentiation track was in line with the developmental-toxic properties of the compounds. The significance of deviation was analyzed using the Leave-One-Out cross validation, which showed a favorable prediction of toxicity in the system. Our findings show that gene-expression signatures can be used to identify developmental-toxicant induced differentiation modulation. In addition, studying compound-induced effects at an early stage of differentiation combined with transcriptomics leads to increased objectivity in determining differentiation inhibition and to a reduction of the test duration. Furthermore, this approach may improve the predictivity and applicability domain of the EST.
    Original languageEnglish
    Pages (from-to)130-139
    Number of pages10
    JournalToxicological Sciences
    Volume116
    Issue number1
    DOIs
    Publication statusPublished - Jul 2010

    Keywords

    • embryonic stem cell
    • differentiation
    • gene expression
    • predictive toxicology
    • microarray
    • embryonic stem cell test
    • VITRO EMBRYOTOXICITY TESTS
    • IN-VITRO
    • THYMIDYLATE SYNTHETASE
    • RAT HEPATOCYTES
    • RETINOIC ACID
    • VALPROIC ACID
    • CARDIOMYOCYTES
    • IDENTIFICATION
    • NORMALIZATION
    • VALIDATION

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