GENETIC VARIATION IN APOLIPOPROTEIN E EXPRESSION

载脂蛋白 E 表达的遗传变异

• 批准号: 6324496
• 负责人: Benjamin Tycko
• 金额: $ 32.23万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6324496
• 关键词: Alzheimer's disease; aging; alleles; apolipoprotein E; cell line; disease /disorder onset; gene expression; gene targeting; genetic enhancer element; genetic markers; genetic models; genetic polymorphism; genetic promoter element; genetically modified animals; genotype; human tissue; laboratory mouse; messenger RNA; postmortem; protein localization; protein structure function; racial /ethnic difference; single strand conformation polymorphism; transfection

Data from genetic epidemiological studies suggest that the involvement of genetic variation at the APOE locus in Alzheimer's disease (AD) susceptibility may not be restricted to the well-studied coding polymorphisms, but may also be influenced by polymorphisms in flanking DNA. In particular, we have recently observed an association of the APOC1 HpaI+ allele with AD in groups of people with the "reference" APOE e3/e3 genotype. In this project we will carry out molecular analyses and experiments to test the hypothesis that APOE flanking sequences influence AD susceptibility is that this effect is through the influence of these polymorphisms on allele-specific APOE mRNA expression. First, to test whether AD susceptibility depends on particular combinations of APOE coding and regulatory polymorphisms located in cis on the chromosome, we will develop a method for determining complete haplotypes of polymorphic markers in the APOE promoter, the APOE coding region and the APOC1 promoter and we will generate haplotype data for AD association analysis in the large multi-ethnic population samples discussed in Project 1. Second, using a transfection assay screen, we will test for the existence of functional sequence polymorphisms in the APOE and APOC1 promoters, we will determine the sequence of functional sequence polymorphisms in the APOE and APOC1 promoters, we will determine the sequence of these polymorphisms in the APOE and APOC1 promoters, we will determine the sequence of these polymorphisms and we will apply them as new markers in the haplotype analyses. Third, we will test for functional significance for the individual polymorphisms and haplotypes in vivo by correlating them with allele-specific APOE mRNA levels in human lymphoblastoid cell lines and liver, kidney and brain tissues. Fourth, using apoCI promoter "knockout" mice, we will test a model in which the APOE and APOC1 engage in enhance-competition as an explanation for the observed association of the APOC1 HpaI+ allele with AD. Using transfections and transgenic mice we will also investigate localization of the putative brain-enhancer of APOE and then search for functional polymorphisms in this region. Results of these studies may reveal a basic mechanism for AD which is accessible to pharmacological intervention and may also shed light on the difference sin apparent AD risk who have been observed in different ethnic groups.

来自遗传流行病学研究的数据表明，在阿尔茨海默病（AD）易感性的APOE基因座的遗传变异的参与可能不限于充分研究的编码多态性，但也可能受到侧翼DNA多态性的影响。特别是，我们最近观察到，在具有“参考”APOE e3/e3基因型的人群中，APOC 1 HpaI+等位基因与AD相关。在这个项目中，我们将进行分子分析和实验，以测试的假设，即APOE侧翼序列影响AD易感性是通过这些多态性对等位基因特异性APOE mRNA表达的影响。首先，为了测试AD易感性是否取决于位于染色体顺式的APOE编码和调节多态性的特定组合，我们将开发一种方法来确定APOE启动子、APOE编码区和APOC 1启动子中多态性标记的完整单倍型，并在项目1中讨论的大型多种族人群样本中生成用于AD关联分析的单倍型数据。第二，使用转染试验筛选，我们将测试APOE和APOC 1启动子中功能序列多态性的存在，我们将确定APOE和APOC 1启动子中功能序列多态性的序列，我们将确定APOE和APOC 1启动子中这些多态性的序列，我们将确定这些多态性的序列，并将它们作为新的标记应用于单倍型分析。第三，我们将通过将个体多态性和单倍型与人类淋巴母细胞系和肝、肾和脑组织中等位基因特异性APOE mRNA水平相关联来测试其在体内的功能意义。第四，使用apoCI启动子“敲除”小鼠，我们将测试APOE和APOC 1参与增强竞争的模型，作为观察到的APOC 1 HpaI+等位基因与AD相关性的解释。使用转染和转基因小鼠，我们也将调查定位的APOE的推定脑增强子，然后在这个地区寻找功能多态性。这些研究的结果可能揭示了AD的基本机制，这是药物干预，也可能揭示了在不同种族群体中观察到的AD风险的差异。