Use of toxicogenomics for identifying genetic markers of pulmonary oedema

Dominique Balharry, Victor Oreffo, Roy Richards

Research output: Contribution to journalArticlepeer-review

Abstract

This study was undertaken primarily to identify genetic markers of oedema and inflammation. Mild pulmonary injury was induced following the instillation of the oedema-producing agent, bleomycin (0.5 units). Oedema was then confirmed by conventional toxicology (lavage protein levels, free cell counts and lung/body weight ratios) and histology 3 days post-bleomycin instillation. The expression profile of 1176 mRNA species was determined for bleomycin-exposed lung (Clontech Atlas macroarray, n=9). To obtain pertinent results from these data, it was necessary to develop a simple, effective method for bioinformatic analysis of altered gene expression. Data were log10 transformed followed by global normalisation. Differential gene expression was accepted if: (a) genes were statistically significant (P < or = 0.05) from a two-tailed t test; (b) genes were consistently outside a two standard deviation (SD) range from control levels. A combination of these techniques identified 31 mRNA transcripts (approximately 3%) which were significantly altered in bleomycin treated tissue. Of these genes, 26 were down-regulated whilst only five were up-regulated. Two distinct clusters were identified, with 17 genes classified as encoding hormone receptors, and nine as encoding ion channels. Both these clusters were consistently down-regulated. The magnitude of the changes in gene expression were quantified and confirmed by Q-PCR (n = 6), validating the macroarray data and the bioinformatic analysis employed. In conclusion, this study has developed a suitable macroarray analysis procedure and provides the basis for a better understanding of the gene expression changes occurring during the early phase of drug-induced pulmonary oedema. This work has been presented orally, in part at the British Association for Lung Research Summer Meeting, University of Brighton, 3-5 September, 2003 and in full at the British Toxicology Society Annual Congress, Heriot Watt University, Edinburgh, 21-24 April 2004.

Original languageEnglish
Pages (from-to)101-8
Number of pages8
JournalToxicology and Applied Pharmacology
Volume204
Issue number2
DOIs
Publication statusPublished - 15 Apr 2005

Keywords

  • Animals
  • Bleomycin
  • Bronchoalveolar Lavage Fluid
  • Down-Regulation
  • Gene Expression Profiling
  • Genetic Markers
  • Inflammation
  • Inflammation Mediators
  • Lung
  • Macrophages, Alveolar
  • Male
  • Multigene Family
  • Organ Size
  • Pulmonary Edema
  • Rats
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction
  • Specific Pathogen-Free Organisms
  • Time Factors
  • Toxicogenetics
  • Transcription, Genetic
  • Up-Regulation
  • Weight Gain

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