1. A systems approach to pathways and networks involved in lipid metabolism in macrophage activation the LIPID MAPS Consortium University Of California San Diego

2. Signaling Pathways Lipid Metabolic Pathways Lipid Associated Proteins Lipid Metabolites Gene expression Protein expression Enzyme activation Cellular Networks

3. Induction of inflammatory and immune response genes Cytokines (IL-1 , TNF  ), Tissue factor, B7, etc. MAP3K (Proteolysis) Macrophage or endothelial cell membrane cell membrane Direct Activation ACTIVATE RAW CELLS WITH Kdo2-Lipid A LIPID MAPS TMTM Measure time series of Gene, Lipid, protein and Cytokine Responses

4. MS Bio Replicates Tech Replicates Mass Spec Runs Layers of Replications and Measurements

5. http://www.lipidmaps.org EXPERIMENTAL DATA

6. Lipidomics Data Display : Gene array-search by pathway http://www.lipidmaps.org/data/results/raw2647/microarray/

7. Lipid Pathways and Networks:Visualization tools VANTED: Displaying lipidomic timecourse data http://vanted.ipk-gatersleben.de/ Hyperlink to protein DB Hyperlink to lipid DB

8. Fatty Acids: Omega-9 polyunsaturated and straight chain fatty acids Integrated lipids, genes, and pathways data across timecourse experiments for RAW 264.7 cells treated with Kdo 2 -lipid A www.lipidmaps.org LIPID MAPS funded by Glue Grant from: www.nigms.nih.gov Fatty Acids: Omega-3 and Omega-6 polyunsaturated fatty acids Fatty Acids: Eicosanoids Sterols Glycerophospholipids Sphingolipids Overview PLA Acetyl CoA carboxylase Malony-CoA decarboxylase Fatty acid synthase Carnitine Palmitoyl Transferase (CPT-1) KDO 2 -lipid A DNA cytosol ACS mitochondria Acyltransferases lipases PLC TLR4 Microarray results Metabolic pathway Signaling pathway Energy metabolism HM G- Co A sy nth as e HM G- Co A red uct as e Serine Palmitoyltransferase Rate limiting step Physical interaction Sterol O-Acyltransferases LCAT LDLHDL ABCA1 ABCG1 Receptors Proteins Lipid rafts PPAR Fatty Acid FAT/CD36FATP Malonyl-CoA Acetyl-CoA HMG-CoA Acyl-CoA Citrate Acyl-CoACitrate Sphingolipids Fatty Acids Glycerolipids Glycerophospholipids SterolsIsoprenoids CE

9. http://www.signalinggateway.org EXPERIMENTAL DATA

10. 2003 Ligand Screen – Time Course cAMP 0.5’ 1’ 3’ 8’ 20’ Phospho Protein 2.5’ 5’ 15’ 30’ Micro Array 30’ 1 hr 2 hr 4 hr Ca++ 10’

11. 2003 Summary of Ligand Screen Responses