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Arch Toxicol. 2017 Jan;91(1):365-391. doi: 10.1007/s00204-016-1690-2. Epub 2016 Mar 25.

Fingerprinting of neurotoxic compounds using a mouse embryonic stem cell dual luminescence reporter assay.

Author information

1
Department of Pathology and Immunology, Medical School, University of Geneva, Geneva, Switzerland.
2
Quretec Ltd, Ülikooli 6a, Tartu, Estonia.
3
Department of Pediatrics, Geneva University Hospital, Geneva, Switzerland.
4
Department of Genetic and Laboratory Medicine, Geneva University Hospital, Centre Medical Universitaire, Rue Michel-Servet, 1211, Geneva 4, Switzerland.
5
NEURIX SA, Geneva, Switzerland.
6
Doerenkamp-Zbinden Chair for In Vitro Toxicology and Biomedicine, University of Konstanz, Constance, Germany.
7
TNO, Zeist, The Netherlands.
8
BioMarin Pharmaceutical Inc., Leiden, The Netherlands.
9
Institute of Neurophysiology and Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.
10
Leibniz Research Centre for Working Environment and Human Factors (IfADo), Technical University of Dortmund, Dortmund, Germany.
11
Department of Pathology and Immunology, Medical School, University of Geneva, Geneva, Switzerland. Karl-Heinz.Krause@unige.ch.
12
Department of Genetic and Laboratory Medicine, Geneva University Hospital, Centre Medical Universitaire, Rue Michel-Servet, 1211, Geneva 4, Switzerland. Karl-Heinz.Krause@unige.ch.

Abstract

Identification of neurotoxic drugs and environmental chemicals is an important challenge. However, only few tools to address this topic are available. The aim of this study was to develop a neurotoxicity/developmental neurotoxicity (DNT) test system, using the pluripotent mouse embryonic stem cell line CGR8 (ESCs). The test system uses ESCs at two differentiation stages: undifferentiated ESCs and ESC-derived neurons. Under each condition, concentration-response curves were obtained for three parameters: activity of the tubulin alpha 1 promoter (typically activated in early neurons), activity of the elongation factor 1 alpha promoter (active in all cells), and total DNA content (proportional to the number of surviving cells). We tested 37 compounds from the ESNATS test battery, which includes polypeptide hormones, environmental pollutants (including methylmercury), and clinically used drugs (including valproic acid and tyrosine kinase inhibitors). Different classes of compounds showed distinct concentration-response profiles. Plotting of the lowest observed adverse effect concentrations (LOAEL) of the neuronal promoter activity against the general promoter activity or against cytotoxicity, allowed the differentiation between neurotoxic/DNT substances and non-neurotoxic controls. Reporter activity responses in neurons were more susceptible to neurotoxic compounds than the reporter activities in ESCs from which they were derived. To relate the effective/toxic concentrations found in our study to relevant in vivo concentrations, we used a reverse pharmacokinetic modeling approach for three exemplary compounds (teriflunomide, geldanamycin, abiraterone). The dual luminescence reporter assay described in this study allows high-throughput, and should be particularly useful for the prioritization of the neurotoxic potential of a large number of compounds.

KEYWORDS:

In vitro screening; Mouse embryonic stem cells; Neuroactivity; Neuronal differentiation; Neurotoxicity

PMID:
27015953
PMCID:
PMC5225183
DOI:
10.1007/s00204-016-1690-2
[Indexed for MEDLINE]
Free PMC Article

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