CCR2 AND CCR5--ATHEROSCLEROSIS AND CONTROL OF EXPRESSION

CCR2 和 CCR5——动脉粥样硬化及其表达控制

• 批准号: 6030684
• 负责人: ISRAEL F. CHARO
• 金额: $ 38.92万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-15 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6030684
• 关键词: apolipoprotein E; atherosclerosis; chemokine; cytokine receptors; gene targeting; genetically modified animals; green fluorescent proteins; heart transplantation; homologous transplantation; immunogenetics; laboratory mouse; molecular cloning; monocyte chemoattractant protein 1; protein structure function; receptor binding; receptor coupling; receptor expression; transplant rejection

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.

描述：（改编自应用程序）此项目的总体目标 提议是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）特异性吸引白细胞 通过激活 G 蛋白偶联受体。 调查人员重点关注 他们对其中两种受体 CCR2 和 CCR5 的努力，这两种受体被克隆在 该实验室和 哪些是单核细胞上的主要趋化因子受体。 在第一个资助期间，研究人员检查了 这两种受体的结构/活性关系 配体结合和信号传导。 当前申请中提出的工作 将利用这些体外研究扩展到体内模型 我们最近创建了 CCR2 敲除小鼠。 该队的第一个目标是 建议确定 CCR2 在两种形式的血管疾病中的作用： 动脉粥样硬化和同种异体心脏加速动脉硬化 移植。 研究人员将 CCR2-/- 小鼠与 apoE-/- 进行杂交 小鼠并进行详细的动脉粥样硬化研究。 调查人员将 还进行同种异体心脏异位移植到 CCR2-/- 小鼠并确定 CCR2 是否在慢性心肌排斥反应中发挥作用 和移植动脉硬化。 第二个目标是创建 CCR2/CCR5 双敲除小鼠。 调查人员将有选择地删除这些 受体并同时在 CCR2 的控制下“敲入”lacZ CCR5 控制下的启动子和绿色荧光蛋白 (GFP) 发起人。 这种方法将产生趋化因子受体敲除动物， 以及提供检测体内内源性的灵敏手段 CCR2 和 CCR5 的表达。 趋化因子受体的差异调节 聚集在 3 号染色体 (3p21.3-24) 上可能提供特异性 炎症和免疫。 第三个目标是确定 组织特异性表达和转录调控（如果选择 CC） 受体。 研究人员将使用半合子 CCR2/CCR5 双敲除小鼠（CCR2+/LacZ、CCR5=/GfP）以及转基因小鼠 用人类基因组 DNA 的大片段创建，以跟踪表达 这两种受体在人类疾病模型中的作用。 提出的实验 在这笔赠款中，将利用新颖和互补的方法来提供 关于趋化因子和趋化因子受体的作用的第一个详细信息 在血管疾病中。