Analysis of cell death inducing compounds.
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.
Spicker
Jeppe S
JS
Center for Biological Sequence Analysis, BioCentrum-DTU, Technical University of Denmark, Kemitorvet, Building 208, 2800, Lyngby, Denmark. hbjorn@cbs.dtu.dk
Pedersen
Henrik Toft
HT
Nielsen
Henrik Bjørn
HB
Brunak
Søren
S
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D018554:Avena / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D016923:Cell Death / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D000072467:Cytochrome P450 Family 2; D020869:Gene Expression Profiling; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D010105:Oxygenases / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley
D002352:Carrier Proteins; C080906:glucokinase regulatory protein; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; C086292:Cyp2c29 protein, mouse; D000072467:Cytochrome P450 Family 2; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase