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Analysis of cell death inducing compounds

  • Molecular Toxicology
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Abstract

Biomarkers for early detection of toxicity hold the promise of improving the failure rates in drug development. In the present study, gene expression levels were measured using full-genome RAE230 version 2 Affymetrix GeneChips on rat liver tissue 48 h after administration of six different compounds, three toxins (ANIT, DMN and NMF) and three non-toxins (Caeruelein, Dinitrophenol and Rosiglitazone). We identified three gene transcripts with exceptional predictive performance towards liver toxicity and/or changes in histopathology. The three genes were: glucokinase regulatory protein (GCKR), ornithine aminotransferase (OAT) and Cytochrome P450, subfamily IIC (mephenytoin 4-hydroxylase) (Cyp2C29). RT-PCR for these three genes was performed and four additional compounds were included for validation. The quantitative RT-PCR analysis confirmed the findings based on the microarray data and using the three genes a classification rate of 55 of 57 samples was achieved for the classification of not toxic versus toxic. The single most promising biomarker (OAT) alone resulted in a surprisingly 100% correctly classified samples. OAT has not previously been linked to toxicity and cell death in the literature and the novel finding represents a putative hepatotoxicity biomarker.

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Abbreviations

Ac:

Acivicin

ANIT:

α-Naftylisothiocyanate

At:

Aflatoxin

BHT:

Butylated hydroxytoluene

BS:

Buthinionine sulphoxide

Ca:

Caeruelein

Con:

Control (untreated) animal

Cyp2C:

Cytochrome P450, family 2, subfamily C

DMF:

Dimethylformamide

DMN:

Dimethylnitrosamine

DNP:

Dinitrophenol

Fmo1:

Flavin containing monooxygenase 1

GCKR:

Glucokinase regulatory protein

KNN:

K-nearest neighbor

NMF:

N-methylformamide

OAT:

Ornithine aminotransferase

Ro:

Rosiglitazone

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Acknowledgments

I would like to thank the COMET I consortium for giving me access to the tissues analyzed in this study. Homepage of the COMET consortium: http://www.bc-comet.sk.med.ic.ac.uk/

I would also like to thank Anne-Marie Mølck from Novo Nordisk with help on toxicity, Nina Hagen for inputs on P450, Lene Normann Nielsen on help with the QPCR and Kristine Dahlin from DTU with assistance in the laboratory.

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Authors and Affiliations

  1. Center for Biological Sequence Analysis, BioCentrum-DTU, Technical University of Denmark, Kemitorvet, Building 208, 2800, Lyngby, Denmark

    Jeppe S. Spicker, Henrik Bjørn Nielsen & Søren Brunak

  2. Protein Structure and Biophysics, Protein Engineering, Novo Nordisk Park, 2760, Måløv, Denmark

    Jeppe S. Spicker & Henrik Toft Pedersen

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  1. Jeppe S. Spicker
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  2. Henrik Toft Pedersen
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Correspondence to Jeppe S. Spicker.

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Spicker, J.S., Pedersen, H.T., Nielsen, H.B. et al. Analysis of cell death inducing compounds. Arch Toxicol 81, 803–811 (2007). https://doi.org/10.1007/s00204-007-0207-4

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  • DOI: https://doi.org/10.1007/s00204-007-0207-4

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