Multiple Agonist Induced Changes in Eicosanoid Metabolites Correlated with Gene Expression in Macrophages

Arachidonic acid is released and converted into hundreds of different bioactive mediators by a variety of cyclooxygenases (COX), lipoxygenases (LO) and cytochrome P450s (CYP). Due to the size and diversity of the eicosanoid class of signaling molecules produced, a thorough and systematic investigation of these biological processes requires the simultaneous quantitation of a large number of eicosanoids in a single analysis. We have developed a LC/MS/MS method that can identify and quantitate over 60 eicosanoids with standards in a single analysis, and applied it to RAW264.7 murine macrophage cells. Changes in the metabolic profile following stimulation with 21 different agonist conditions were correlated to gene expression using microarray analysis, showing the interplay between genomics and metabolomics. This approach allows us to investigate potential novel eicosanoid signaling networks with an array of different agonists and correlate them with gene induction and metabolite profiles. This work was supported by the LIPID MAPS Large Scale CollaborativebGrant GM-069338 from the National Institutes of Health.