Genonic analysis to Identify atherosclerosis-susceptible and -resistant genes and Investigation of molecular mechanism In the pathogenesis of atherosclerosis

本项目的具体目的是：(1)通过分析动脉粥样硬化易感和耐药小鼠，鉴定动脉粥样硬化易感和耐药基因；(2)利用候选基因的单核苷酸多态性（snp）和新发现的动脉粥样硬化易感和耐药基因进行遗传分析。项目进展情况如下：1；为了鉴定动脉粥样硬化易感基因和耐药基因，分别饲喂高胆固醇饮食的动脉粥样硬化易感小鼠（C57BL/6J）和动脉粥样硬化耐药小鼠（C3H/HeJ）。随后，从这些动物的血管内皮细胞、血管平滑肌细胞和骨髓干细胞中提取mrna，用cDNA微阵列芯片进行表达谱分析（In viva expression profiling）。进行了表达谱分析，目前正在分析收集到的数据。同时，从这些动物的血管内皮细胞、血管平滑肌细胞和骨髓干细胞中提取更多的培养细胞，进行体外表达谱分析。在几个候选基因中，确定MCP-1 A-2518G多态性和CCR2 G270A多态性的实验条件已经确定。然后，对候选基因如MCP-1、TGF-β和NF-κB进行多态性搜索，但未发现改变基因功能的新多态性。同时，继续招募缺血性心脏病（IHD）患者，但在300例IHD患者中仅发现2例。我们重新分析了患者数据库，发现许多冠状动脉危险因素对动脉粥样硬化的发病机制影响很小。因此，我们改变了患者招募策略，开始招募符合新标准的IHD患者。未来，将利用动脉粥样硬化相关候选基因的多态性进行遗传分析。少

Specific aims of this project are (1)to Identify atherosclerosis-susceptible and -resistant genes by analyzing atherosclerosis-susceptible and -resistant mice, and (2)to carry out genetic analysis by using single nucleotide polymorphisms (SNPs) of candidate genes and newly identified atherosclerosis-susceptible and -resistant genes. Progress of this project is as follows;1. Identification of atherosclerosis-susceptible and -resistant genesTo identity atherosclerosis-susceptible and -resistant genes, atherosclerosis-susceptible mouse line (C57BL/6J) and atherosclerosis -resistant mouse line (C3H/HeJ), were fed with high cholesterol diet. Subsequently, mRNAs were extracted from vascular endothelial cells, vascular smooth muscle cells and bone marrow stem cells of these animals for expression profiling (In viva expression profiling) with cDNA microarray chips. Expression profiling was carried out, and collected data are now under analysis. At the same time, optimization for generating pri … More mary cultured cells from vascular endothelial cells, vascular smooth muscle cells and bone marrow stem cells of these animals are undergone to perform in vitro expression profiling.2.Identification of sequence variations in atherosclerosis-related candidate genes for genetic analysisAmong several candidate genes experimental condition to determine MCP-1 A-2518G polymorphism and CCR2 G270A polymorphism has been set. Then, polymorphisms of candidate genes, such as MCP-1,TGF-β and NF-κB, were searchd, but no novel polymorphisms to alter gene function were identified. At the same time, recruitment of ischemic heart diseasee(IHD)patients has been continued,but only 2 out of 300 IHD patients were found. Our patient database was reanalyzed, and It was identified that number of coronary risk factors had an small influence on the pathogenesis of atherosclerosis. Therefore, a strategy of patient recruitment was changed, and recruitment of IHD patients with new criteria was begun. ln future, genetic analysis using polymorphisms of aterosclerosis-related candidate genes will be conducted. Less