Fractalkine: Roles in Cell Adhesion and Atherosclerosis

描述（由申请人提供）：趋化因子在血管疾病中单核细胞和巨噬细胞的运输中起关键作用，但对其作用的分子机制知之甚少。Fractalkine是一种具有不寻常结构的新型趋化因子。与可溶性趋化因子不同，fractalkine由与膜结合的粘蛋白柄融合的趋化因子样结构域组成。通过TACE的作用产生可溶形式的fractalkine，TACE是一种金属蛋白酶，其刚好在跨膜结构域上方切割fractalkine。在其可溶形式中，fractalkine是一种有效的化学引诱物，但在其膜结合形式中，它捕获携带其同源受体CX 3CR 1的细胞。我们已经证明，CX 3CR 1通过基因靶向缺失的小鼠可以防止异位心脏移植排斥反应和饮食诱导的动脉粥样硬化。然而，这些血管疾病的膜结合和可溶性形式的fractalkine的相对贡献是未知的。为了研究膜结合FK的作用，我们将创建表达不可切割形式的fractalkine的敲入小鼠。为了研究可溶性fractalkine的作用，我们将创建表达可溶性但不表达膜结合FK的小鼠。这些研究将检验FK的裂解有助于动脉粥样硬化形成的假设。最近对患者的遗传学研究表明，CX 3CR 1的多态性与冠状动脉疾病的保护相关。我们将把这种V259 I/T280 M突变引入小鼠CX 3CR 1中，并测试该多态性通过防止单核细胞/巨噬细胞的捕获而提供对动脉粥样硬化的保护的假设。Fractalkine并不是唯一被发现在动脉粥样硬化中发挥作用的趋化因子。我们以前已经表明，MCP-1，及其受体CCR 2，有助于巨噬细胞的招聘和病变的形成。在本提案的最后部分，我们将创建缺乏fractalkine和CCR 2的双敲除小鼠，以确定这些趋化因子是否独立或协同作用以促进动脉粥样硬化。这些目标的完成将大大推进我们对fractalkine促进动脉粥样硬化病变形成的机制的理解。

DESCRIPTION (provided by applicant): Chemokines play a critical role in the trafficking of monocytes and macrophages in vascular disease, but the molecular mechanisms of their actions are poorly understood. Fractalkine is a novel chemokine with an unusual architecture. Unlike soluble chemokines, fractalkine consists of a chemokine-like domain fused to a membrane-bound mucin stalk. A soluble form of fractalkine is created by the action of TACE, a metalloprotease that cleaves fractalkine just above the transmembrane domain. In its soluble form fractalkine is a potent chemoattractant, but in its membrane-bound form it captures cells bearing its cognate receptor, CX3CR1. We have shown that mice in which CX3CR1 is deleted by gene targeting are protected against rejection of heterotopic cardiac transplants, and diet-induced atherosclerosis. However, the relative contributions of the membrane-bound and soluble forms of fractalkine to these vascular disease are unknown. To investigate the role of membrane-bound FK we will create knock-in mice expressing a noncleavable form of fractalkine. To investigate the role of soluble fractalkine, we will create mice that express soluble, but not membrane-bound FK. These studies will test the hypothesis that the cleavage of FK contributes to atherogenesis. Recent genetic studies in patients have revealed that a polymorphism in CX3CR1 correlates with protection from coronary artery disease. We will introduce this V259I/T280M mutation into murine CX3CR1, and test the hypothesis that the polymorphism affords protection from atherosclerosis by preventing the capture of monocyte/macrophages. Fractalkine is not the only chemokine that has been found to play a role in atherosclerosis. We have previously shown that MCP-1, and its receptor CCR2, contribute to macrophage recruitment and lesion formation. In the final portion of this proposal, we will create double knockout mice lacking both fractalkine and CCR2 to determine if these chemokines act independently or in concert to promote atherogenesis. Completion of these aims will significantly advance our understanding of the mechanisms by which fractalkine contributes to lesion formation in atherosclerosis.