CCR2 AND CCR5--ATHEROSCLEROSIS AND CONTROL OF EXPRESSION

描述：(改编自应用程序)这项工作的总体目标 建议确定趋化因子受体在血管中的作用 疾病。趋化因子(趋化细胞因子)特异性地吸引白细胞 通过激活G蛋白偶联受体。调查人员已经将注意力集中在 他们在其中两个受体CCR2和CCR5上的努力，这两个受体被克隆到 这个实验室和单核细胞上的主要趋化因子受体。 在第一个资助期内，调查人员审查了 这两种受体的结构/活性关系 配体结合和信号转导。在当前申请中提出的工作 将把这些体外研究扩展到体内模型，利用 我们最近创造了一种CCR2基因敲除小鼠。第一个目标是 建议确定CCR2在两种形式的血管疾病中的作用： 动脉粥样硬化与同种异体心脏动脉硬化 移植。研究人员将与携带apoE-/-的CCR2-/-小鼠进行杂交 并进行详细的动脉粥样硬化研究。调查人员将 也进行异体心脏到CCR2的异位移植-/- 并确定CCR2是否在慢性心肌排斥反应中起作用 和移植动脉硬化。第二个目标是创建CCR2/CCR5 双基因敲除小鼠。调查人员将有选择地删除这些 受体与CCR2控制下的LacZ同时“敲入” CCR5控制下的启动子和绿色荧光蛋白 推动者。这种方法将产生趋化因子受体基因敲除动物， 并提供了一种检测体内内源性的敏感手段 CCR2和CCR5的表达。趋化因子受体的差异性调节 聚集在3号染色体(3p21.3-24)上可能提供特异性 炎症和免疫力。第三个目标是确定 如果选择CC，则组织特异性表达和转录调控 感受器。研究人员将使用半合子CCR2/CCR5 双基因敲除小鼠(CCR2+/LacZ，CCR5=/GFP)以及转基因小鼠 用人类基因组DNA的大片段创建，以遵循表达 这两种受体在人类疾病的模型中。建议进行的实验 在这笔赠款中，将利用新颖和互补的方法提供 趋化因子和趋化因子受体作用的第一个详细信息 在血管疾病方面。

DESCRIPTION: (Adapted from the application) The overall goal of this proposal is to determine the role of chemokine receptors in vascular disease. Chemokines (chemotatic cytokines) specifically attract leukocytes by activating G-protein-coupled receptors. The investigators have focused their efforts on two of these receptors, CCR2 and CCR5, which were cloned in this laboratory and which are the major chemokine receptors on monocytes. During the first funding period, the investigators examined the structure/activity relationships of these two receptors with regard to ligand binding and signaling. The work proposed in the current application will extend these in vitro studies to in vivo models, taking advantage of our recent creation of a CCR2 knockout mouse. The first goal of the proposal is to determine the role of CCR2 in two forms of vascular disease: atherosclerosis and the accelerated arteriosclerosis of allogeneic cardiac transplants. The investigators will cross the CCR2-/- mice with apoE-/- mice and perform detailed atherosclerosis studies. The investigators will also perform heterotopic transplants of allogeneic hearts into the CCR2-/- mice and determine whether CCR2 plays a role in chronic myocardial rejection and transplant arteriosclerosis. The second goal is creating CCR2/CCR5 double-knockout mice. The investigators will selectively delete these receptors and simultaneously "knock in" lacZ under the control of the CCR2 promoter and green fluorescent protein (GFP) under the control of the CCR5 promoter. This approach will generate chemokine receptor knockout animals, as well as provide a sensitive means for detecting the endogenous in vivo expression of CCR2 and CCR5. Differential regulation of chemokine receptors clustered on chromosome 3 (3p21.3-24) may provide specificity in inflammation and immunity. The third goal is to determine the tissue-specific expression and transcriptional regulation if selected CC receptors. The investigators will use the hemizygous CCR2/CCR5 double-knockout mice (CCR2+/LacZ, CCR5=/GfP), as well as transgenic mice created with large fragments of human genomic DNA, to follow the expression of these two receptors in models of human disease. the experiments proposed in this grant will utilize novel and complementary approaches to provide the first detailed information on the role of chemokines and chemokine receptors in vascular disease.