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

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

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

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

http://www.lipidmaps.org EXPERIMENTAL DATA

Lipidomics Data Display : Gene array-search by pathway http://www

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

Integrated lipids, genes, and pathways data across timecourse experiments for RAW 264.7 cells treated with Kdo2-lipid A LIPID MAPS funded by Glue Grant from: www.nigms.nih.gov www.lipidmaps.org Overview PLA carboxylase Acetyl CoA Malony-CoA decarboxylase synthase Fatty acid Carnitine Palmitoyl Transferase (CPT-1) KDO2-lipid A DNA cytosol ACS mitochondria Acyltransferases lipases PLC TLR4 Microarray results Metabolic pathway Signaling pathway Energy metabolism HMG-CoA reductase Serine Palmitoyltransferase Rate limiting step Physical interaction Sterol O-Acyltransferases LCAT LDL HDL ABCG1 ABCA1 Receptors Proteins Lipid rafts PPAR Fatty Acid FAT/CD36 FATP Malonyl-CoA Acetyl-CoA Acyl-CoA Citrate Sphingolipids Fatty Acids Glycerolipids Glycerophospholipids Sterols Isoprenoids CE Fatty Acids: Omega-3 and Omega-6 polyunsaturated fatty acids Sterols Glycerophospholipids Fatty Acids: Eicosanoids Fatty Acids: Omega-9 polyunsaturated and straight chain fatty acids Sphingolipids

http://www.signalinggateway.org EXPERIMENTAL DATA

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

Summary of Ligand Screen Responses