Abstract 117: AIBP Deficiency Affects Inflammarafts Formation And Macrophage Reprogramming In Atherosclerosis

Background: Excessive accumulation of cholesterol in plasma membrane of vascular macrophages leads to formation of inflammarafts - large, cholesterol-rich lipid rafts hosting inflammatory receptors assemblies - and reprograms macrophages to facilitate hyperinflammatory responses. Removal of cholesterol from membrane and storage in intracellular lipid droplets reduces macrophage inflammatory status. Apolipoprotein A-I binding protein (AIBP) has been found to mediate selective cholesterol efflux and lipid raft dynamics regulation via binding to toll-like receptor-4 (TLR4) thus reducing inflammarafts in activated macrophages. However, the underlying mechanisms driving lipid raft dynamics changes of reprogrammed macrophages remain unclear. The objective of this study was to evaluate the effect of AIBP deficiency on inflammaraft formation and macrophage reprogramming, separately in macrophage foam cells and non-foamy macrophages. Methods: Single-cell suspensions from the aortae of 16-week high-fat diet-fed Apoa1bp -/- Ldlr -/- and Ldlr -/- mice were gated for BODIPY-high foamy and BODIPY-low non-foamy CD45 + F4/80 + macrophages by flow cytometry. Lipid droplets were assessed by BODIPY fluorescence and inflammarafts were characterized by increased levels of lipid rafts and TLR4 dimers. Transcriptome and chromatin accessibility were measured by RNA-seq and ATAC-seq. Plasma lipid levels and size of atherosclerotic lesions and necrotic cores at the aortic origin were measured using standard methods. Results: We found a larger number of foamy macrophages in Apoa1bp -/- Ldlr -/- mice, expressing significantly higher levels of lipid rafts and TLR4 dimers than those in Ldlr -/- mice. However, there were no inflammarafts differences between non-foamy macrophages in Apoa1bp -/- Ldlr -/- and Ldlr -/- mice. The inflammarafts measures were associated with plasma cholesterol levels and size of atherosclerotic lesions and necrotic cores. Inflammatory pathways were up-regulated in Apoa1bp -/- Ldlr -/- foamy macrophages, compared to Ldlr -/- mice. Conclusions: Our results suggest that AIBP helps reduce inflammarafts specifically in macrophage foam cells, rendering them less inflammatory as shown by transcriptomic and open chromatin analyses.