APP GENE PROMOTER IN ALZHEIMER'S DISEASE

描述：（申请人摘要中的逐字记录） B-淀粉样前体蛋白（APP）基因的转录与 阿尔茨海默病（AD）的发病机制。一些早期的研究表明， AD脑组织APP基因表达可能增加。这一增长在 APP表达可导致AD，其在AD患者中的高发病率强烈提示APP表达可导致AD。 唐氏综合症患者有三个拷贝的基因。AD是一种复杂的 可能有多种诱因的疾病很可能 APP转录调节的紊乱影响AD的一个亚类 患者或者，未能持续观察到可检测的增加 死后大脑中APP mRNA的表达可能是由于 APP，这可能导致淀粉样斑块的成核。我们的假设是 APP调节通路在AD发病机制中起关键作用。我们 长期目标是研究APP基因的转录调控。我们有 最近克隆并鉴定了7.9kb的APP启动子区。这里我们将 研究APP启动子上游调控元件（URE）的作用 活动和迟发性AD。这项提议是基于我们的新发现， 一个-75到+104 bp的区域在细胞类型特异性表达中调节APP启动子活性。 方式具体目的是：1）确定APP启动子的核心区 这对它的活动和不朽性至关重要。某些上游 在神经元中赋予基本启动子活性的调节区， 星形胶质细胞可能对生长因子和细胞因子的激活有反应， 在AD的炎症反应期间由活化的巨噬细胞产生。 调控区的功能表征将通过一系列的 细胞系和原代培养物中的缺失策略和DNA转染。（二） 确定URE是否与细胞类型特异性核蛋白相互作用。我们 将研究特定的上游区域如何与细胞类型特异性 神经元和星形胶质细胞中的因子，以及这种相互作用将如何在 细胞因子和生长因子的存在。3)确定统一人力资源管理局的作用 在发育和疾病状态下。启动子的差异效应 细胞类型中的区域可能表明URE在发育过程中的作用， 分化我们计划检查尿素结合因子的水平， 发育大鼠脑，正常和AD脑。4)为了确定基因 AD中调控区的变异性。遗传变异性 调节区的表达对于迟发性AD散发受试者可能是重要的。我们 建议筛选基因组DNA的“启动子元件”，并比较 “启动子结合因子”在对照组和AD受试者中的表达。5)操纵 启动子活性。我们计划修改具体的监管规定 通过体内竞争实验和使用 反义寡核苷酸策略。最后，我们将启动子 对照组和AD受试者中APP和AB水平的研究。了解 启动子结构域和转录因子之间复杂的相互作用 可能为治疗干预提供新的方法。

DESCRIPTION: (Verbatim from the Applicant's Abstract) Misregulation of transcription of theB-amyloid precursor protein (APP) gene is implicated in the Pathogenesis of Alzheimer's disease (AD). Several early studies indicated that the APP gene expression might be increased in AD brains. That increases in the APP expression can lead to AD is strongly suggested by its high incidence in Down's syndrome patients with three copies of the gene. AD is a complex disorder with potentially multiple triggers. It is quite possible that disturbance in the transcriptional regulation of APP affects a subset of AD patients. Alternatively, failure to observe consistently a detectable increase in APP mRNA in post-mortem brains may be due to a transient over expression of APP, which may lead to nucleation of amyloid plaques. Our hypothesis is that the APP regulatory pathways play a critical role in AD pathogenesis. Our long-term goal is to study the transcriptional control of the APP gene. We have recently cloned and characterized a 7.9 kb APP promoter region. Here we will examine the role of the upstream regulatory elements (URE) on APP promoter activity and in late-onset AD. This proposal is based on our novel finding that a -75 to +104 bp region regulates APP promoter activity in a cell-type specific manner. The specific aims are: 1) To determine the coredomain of APP promoter that is essential for its activity and indelibility. Certain upstream regulatory regions that confer basic promoter activity in neurons and astrocytes may respond to activation by growth factors and cytokines, which are produced by activated macrophages during the inflammatory response in AD. Functional characterization of the regulatory regions will be done by a serial deletion strategy and DNA transfection in cell lines and primary cultures. 2) To identify whether URE interacts with a cell type specific nuclear protein. We will study how the specific upstream region interacts with a cell type-specific factor in neurons and astrocytes and how this interaction will be affected in the presence of cytokines and growth factors. 3) To determine the role of URE in the developmental and disease state. Differential effects of the promoter region in cell types might suggest a role for URE during development and differentiation. We plan to examine the levels of URE-binding factor in developmental rat brains, normal and AD brains. 4) To determine the genetic variability of the regulatory region in AD. The genetic variability of the regulatory region may be important for late-onset AD sporadic subjects. We propose to screen genomic DNA for 'promoter elements' and to compare the 'promoter binding factors' in control and AD subjects. 5) To manipulate promoter activity in cell culture. We plan to modify the specific regulatory effect of the APP promoter by in vivo competition experiment and using the antisense oligodeoxynucleotide strategy. Finally, we will correlate promoter studies with levels of APP and AB in control and AD subjects. Understanding the complex interplay between the promoter domains and the transcription factors may suggest novel methods for therapeutic intervention.