Concentration-dependent gene expression responses to flusilazole in embryonic stem cell differentiation cultures

Journal Article · Tue Mar 01 04:00:00 EST 2011 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2010.12.008· OSTI ID:21535250

Pennings, Jeroen L.A. ^[1]; Netherlands Toxicogenomics Centre, Maastricht ^[2]; Brauers, Karen J.J.; Claessen, Sandra ^[3]; Delft, Joost H. van; Kleinjans, Jos C.S. ^[3]; Netherlands Toxicogenomics Centre, Maastricht ^[2]; Netherlands Toxicogenomics Centre, Maastricht ^[2]

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1. Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands)
2. Netherlands
3. Department of Health Risk Analysis and Toxicology (GRAT), Nutrition and Toxicology Research Institute Maastricht (NUTRIM), Maastricht University, Maastricht (Netherlands)

The murine embryonic stem cell test (EST) is designed to evaluate developmental toxicity based on compound-induced inhibition of embryonic stem cell (ESC) differentiation into cardiomyocytes. The addition of transcriptomic evaluation within the EST may result in enhanced predictability and improved characterization of the applicability domain, therefore improving usage of the EST for regulatory testing strategies. Transcriptomic analyses assessing factors critical for risk assessment (i.e. dose) are needed to determine the value of transcriptomic evaluation in the EST. Here, using the developmentally toxic compound, flusilazole, we investigated the effect of compound concentration on gene expression regulation and toxicity prediction in ESC differentiation cultures. Cultures were exposed for 24 h to multiple concentrations of flusilazole (0.54-54 {mu}M) and RNA was isolated. In addition, we sampled control cultures 0, 24, and 48 h to evaluate the transcriptomic status of the cultures across differentiation. Transcriptomic profiling identified a higher sensitivity of development-related processes as compared to cell division-related processes in flusilazole-exposed differentiation cultures. Furthermore, the sterol synthesis-related mode of action of flusilazole toxicity was detected. Principal component analysis using gene sets related to normal ESC differentiation was used to describe the dynamics of ESC differentiation, defined as the 'differentiation track'. The concentration-dependent effects on development were reflected in the significance of deviation of flusilazole-exposed cultures from this transcriptomic-based differentiation track. Thus, the detection of developmental toxicity in EST using transcriptomics was shown to be compound concentration-dependent. This study provides further insight into the possible application of transcriptomics in the EST as an improved alternative model system for developmental toxicity testing.

• OSTI ID: 21535250
• Journal Information: Toxicology and Applied Pharmacology, Journal Name: Toxicology and Applied Pharmacology Journal Issue: 2 Vol. 251; ISSN TXAPA9; ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ANIMAL CELLS
BIOSYNTHESIS
DETECTION
EVALUATION
FORECASTING
GENES
INHIBITION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RISK ASSESSMENT
RNA
SENSITIVITY
SOMATIC CELLS
STEM CELLS
SYNTHESIS
TESTING
TOXICITY