Although tremendous progress has been made in primary and secondary prevention, the complications of atherosclerosis (heart attacks, strokes and peripheral artery disease) still account for the most deaths worldwide. Here, we propose to test three hypotheses. This work has become possible through the intravital live cell triggered imaging system (ILTIS) we developed in the current funding cycle. Hypothesis 1 is that non- classical monocytes use LFA-1 (?L?2 integrin) and ~40% also VLA-4 (?4?1 integrin) to patrol the endothelium overlying atherosclerotic plaque. We have preliminary evidence for at least two subsets of patrolling monocytes in atherosclerotic arteries. To test the molecular mechanism of patrolling, we will block integrins, their ligands, chemokines and chemokine receptors using monoclonal antibodies. Hypothesis 2 is that blood monocytes gain access to atherosclerotic plaque through the luminal endothelium. To test how monocytes enter atherosclerotic lesions, we have made mice with yellow (YFP) endothelium that are crossed into the Apoe-/- atherosclerosis- prone background. Combining these mice with mice in which monocyte subsets are fluorescently labeled (Cx3cr1GFP/+ for non-classical, Ccr2RFP/+ for classical monocytes) will show transendothelial migration of monocytes into atherosclerotic plaque in vivo. Hypothesis 3 is that four newly discovered plaque macrophage subsets (by ILTIS) have different functions. Preliminary RNA-Seq data show that their gene expression profiles vary significantly. We will test these functions: proliferation by BrdU and Ki67; oxLDL and minimally modified (mm)LDL uptake; efferocytosis; migration and antigen presentation. When the proposed work is complete, we will know, for the first time, how monocytes patrol atherosclerotic arteries, how they enter atherosclerotic lesions and what they do there. We will identify key functions of the four newly discovered monocyte and macrophage subsets in atherosclerotic plaque.
Although tremendous progress has been made in primary and secondary prevention, the complications of atherosclerosis (heart attacks, strokes and peripheral artery disease) still account for the most deaths worldwide. Atherosclerotic plaques are full of macrophages, a type of white blood cell that causes inflammation. We have developed a unique microscope and image processing tools that can look at atherosclerotic plaque in live mice in which these cells are labeled by genetically encoded colors. We see white blood cells patrolling the inside of the artery at the spot overlying the atherosclerotic plaque. Using modern deep sequencing technology, we have discovered significantly different genetic programs in these cells. We propose to test what the functions of each of these cell types are.
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|Winkels, Holger; Ehinger, Erik; Vassallo, Melanie et al. (2018) Atlas of the Immune Cell Repertoire in Mouse Atherosclerosis Defined by Single-Cell RNA-Sequencing and Mass Cytometry. Circ Res 122:1675-1688|
|Anto Michel, Nathaly; Colberg, Christian; Buscher, Konrad et al. (2018) Inflammatory Pathways Regulated by Tumor Necrosis Receptor-Associated Factor 1 Protect From Metabolic Consequences in Diet-Induced Obesity. Circ Res 122:693-700|
|Winkels, Holger; Ley, Klaus (2018) Natural Killer Cells at Ease: Atherosclerosis Is Not Affected by Genetic Depletion or Hyperactivation of Natural Killer Cells. Circ Res 122:6-7|
|Mauler, Maximilian; Herr, Nadine; Schoenichen, Claudia et al. (2018) Platelet Serotonin Aggravates Myocardial Ischemia/Reperfusion Injury via Neutrophil Degranulation. Circulation :|
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|Thiriot, Aude; Perdomo, Carolina; Cheng, Guiying et al. (2017) Differential DARC/ACKR1 expression distinguishes venular from non-venular endothelial cells in murine tissues. BMC Biol 15:45|
|Ley, Klaus; Gerdes, Norbert; Winkels, Holger (2017) ATVB Distinguished Scientist Award: How Costimulatory and Coinhibitory Pathways Shape Atherosclerosis. Arterioscler Thromb Vasc Biol 37:764-777|
|Abadier, Michael; Pramod, Akula Bala; McArdle, Sara et al. (2017) Effector and Regulatory T Cells Roll at High Shear Stress by Inducible Tether and Sling Formation. Cell Rep 21:3885-3899|
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