Myeloid cell interactions in atherosclerosis

描述 尽管在一级和二级预防方面取得了巨大进展，但 动脉粥样硬化(心脏病、中风和外周动脉疾病)仍然是死亡人数最多的原因。 全世界。在这里，我们建议检验三个假设。这项工作已经成为可能通过生命中的 我们在当前资金周期中开发的活细胞触发成像系统(ILTIS)。假设1是非- 经典的单核细胞使用LFA-1(αLβ2整合素)和~40%的VLA-4(α4β1整合素)来巡视内皮 上面有动脉粥样硬化斑块。我们有初步证据表明至少有两个巡逻单核细胞亚群 在动脉粥样硬化的动脉中。为了测试巡逻的分子机制，我们将阻断整合素及其配体， 使用单抗检测趋化因子和趋化因子受体。假设2是血液单核细胞获得 通过管腔内皮细胞进入动脉粥样硬化斑块。测试单核细胞如何进入动脉粥样硬化 损伤，我们已经使小鼠的黄色(YFP)内皮细胞进入APOE-/-动脉粥样硬化- 俯卧背景。将这些小鼠与单核细胞亚群被荧光标记的小鼠相结合 (Cx3cr1GFP/+用于非经典单核细胞，Ccr2RFP/+用于经典单核细胞)将显示 体内单核细胞转化为动脉粥样硬化斑块。假设3是四个新发现的斑块巨噬细胞 子集(由ILTIS提供)具有不同的功能。初步的RNA-Seq数据显示，它们的基因表达谱 差异很大。我们将测试这些功能：BrdU和Ki67的增殖；oxLDL和最低限度的修饰 (Mm)低密度脂蛋白摄取；胞吐作用；迁移和抗原提呈。当建议的工作完成后，我们 将第一次知道单核细胞如何巡视动脉粥样硬化的动脉，如何进入动脉粥样硬化 损伤以及它们在那里做了什么。我们将确定四个新发现的单核细胞的关键功能，并 动脉粥样硬化斑块中巨噬细胞亚群。

Description 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 (αLβ2 integrin) and ~40% also VLA-4 (α4β1 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.