Clofibrate-induced gene expression changes in rat liver: a cross-laboratory analysis using membrane cDNA arrays.
Microarrays have the potential to significantly impact our ability to identify toxic hazards by the identification of mechanistically relevant markers of toxicity. To be useful for risk assessment, however, microarray data must be challenged to determine reliability and interlaboratory reproducibility. As part of a series of studies conducted by the International Life Sciences Institute Health and Environmental Science Institute Technical Committee on the Application of Genomics to Mechanism-Based Risk Assessment, the biological response in rats to the hepatotoxin clofibrate was investigated. Animals were treated with high (250 mg/kg/day) or low (25 mg/kg/day) doses for 1, 3, or 7 days in two laboratories. Clinical chemistry parameters were measured, livers removed for histopathological assessment, and gene expression analysis was conducted using cDNA arrays. Expression changes in genes involved in fatty acid metabolism (e.g., acyl-CoA oxidase), cell proliferation (e.g., topoisomerase II-Alpha), and fatty acid oxidation (e.g., cytochrome P450 4A1), consistent with the mechanism of clofibrate hepatotoxicity, were detected. Observed differences in gene expression levels correlated with the level of biological response induced in the two in vivo studies. Generally, there was a high level of concordance between the gene expression profiles generated from pooled and individual RNA samples. Quantitative real-time polymerase chain reaction was used to confirm modulations for a number of peroxisome proliferator marker genes. Though the results indicate some variability in the quantitative nature of the microarray data, this appears due largely to differences in experimental and data analysis procedures used within each laboratory. In summary, this study demonstrates the potential for gene expression profiling to identify toxic hazards by the identification of mechanistically relevant markers of toxicity.
Baker
Valerie A
VA
Safety and Environmental Assurance Centre, Unilever Research Colworth, Sharnbrook, Bedfordshire, United Kingdom. val.baker@sanofi-synthelabo.com
Harries
Helen M
HM
Waring
Jeff F
JF
Duggan
Colette M
CM
Ni
Hong A
HA
Jolly
Robert A
RA
Yoon
Lawrence W
LW
De Souza
Angus T
AT
Schmid
Judith E
JE
Brown
Roger H
RH
Ulrich
Roger G
RG
Rockett
John C
JC
D000818:Animals; D002994:Clofibrate / Q000633:toxicity*; D020869:Gene Expression Profiling; D000960:Hypolipidemic Agents / Q000633:toxicity*; D008099:Liver / Q000187:drug effects* / Q000473:pathology*; D008297:Male; D015588:Observer Variation; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D018570:Risk Assessment
D000960:Hypolipidemic Agents; D012313:RNA; D002994:Clofibrate