Generation of standards for structural and quantitative assays of novel lipids generated by innate immune cells in humans (360G-Wellcome-094143_Z_10_B)

This grant will focus on extending our understanding of the biological actions and potential therapeutic applications of recently-described hydroxyeicosatetraenoic acid-phospholipids (HETE-PLs), generated by platelets, neutrophils and monocytes/macrophages. A multidisciplinary approach combining biophysics, chemistry, genetics, cell biology and in vivo techniques will be utilized. Basic biochemical studies will be complemented by in vivo murine studies, and clinical studies in a human disorder with known elevated risk of thrombosis and plasma lipid peroxidation. A series of integrated aims is proposed: 1. Determine their role in regulating membrane biophysical behaviour (lysosome formation, phagolysosome fusing, microvesicle formation, etc). 2. Determine their role in regulating function of membrane proteins that require lipid microdomains, e.g. TLRs and their accessory proteins. 3. Determine how they regulate activity of coagulation enzymes at the cell surface. 4. Determine th eir function as ligands for innate immune receptors. 5. Determine how the regulate complement binding to activated immune cells. 6. Determine their role in the pathogenesis of antiphospholipid syndrome in human patients.