INTERDISCIPLINARY APPROACH TO ALZHEIMER DRUG DISCOVERY

该计划的总体目标是应用生物化学，细胞生物学和转基因方法来阐明与阿尔茨海默病（AD）相关的脑载脂蛋白，特别是载脂蛋白E（APOE，基因; apoE，蛋白质）和载脂蛋白J（apoJ）的重要神经生物学活性，并阐明β-淀粉样蛋白生成是如何由这些蛋白质调节的。由于APOE ε 4相关的AD是AD的最普遍形式，我们计划将重点放在确定apoE依赖（特别是ε 4亚型依赖）和apoJ依赖的现象，可能成为治疗目标。在项目1中，研究者将在不存在或存在非变性apoE β 2、ε 3或ε 4 [ROS]、apoJ等的情况下测量apoE-同种型/Abeta复合物形成和Abeta纤维形成。其被认为有助于体内β-淀粉样蛋白生成。这些现象的作用将在其以各种物理状态（可溶的、聚集的/原纤维化的、氧化的）之一供应给原代脑培养物或在apoE ε 2、ε 3或ε 4同种型或apoJ（其反过来将外源地或通过转基因供应）不存在或存在的情况下供应给体内脑后确定。在项目2中，从正常受试者、老年个体和AD患者获得循环可溶性水平。还将确定脂蛋白颗粒中sAbeta分布和apoE亚型对sAbeta分布的影响。将研究非氧化和氧化载脂蛋白。在相关目标中，将使用新的和已证实的策略创建循环sAbeta项目3的清除和降解的细胞位点，脑淀粉样血管病模型。此外，使用含有FAD K670 N/M671 L突变的人β APP cDNA，由鼠Thy-1启动子或ARPP-21神经元特异性核心启动子驱动，构建在神经元细胞中过表达人β APP基因的转基因小鼠。最后，将产生使用神经胶质酸性蛋白启动子在神经胶质细胞中过表达人apoL蛋白的转基因小鼠，并用于确定apoJ在Abeta溶解中的作用。这些项目将由一个行政核心，一个历史学核心和一个Abeta核心支持。“Abeta核心”将监督Abeta结构状态的标准化和Abeta水平的专门、高灵敏度测量。

The overall goal of the Program is to apply biochemical, cell biological, and transgenic and approaches to elucidate the important neurobiological activities of brain apolipoproteins relevant to Alzheimer disease (AD), especially apolipoprotein E (APOE, gene; apoE, protein) and apolipoprotein J (apoJ), and to clarify how beta-amyloidogenesis is regulated by these proteins. Since APOE epsilon4-related AD is the most prevalent form of AD, we plan to focus on identifying apoE-dependent (especially epsilon4-isoform-dependent) and apoJ-dependent phenomena which could become therapeutic targets. In Project 1, investigators will measure apoE-isoform/Abeta complex formation and Abeta fibrillogenesis in the absence or presence of non-denatured apoE epsilon 2, epsilon3 or epsilon4 [ROS], apoJ, ETC.) which are believed to contribute to beta-amyloidogenesis in vivo. The role of these phenomena will be determined after it is supplied in one of various physical states (soluble, aggregated/fibrilized, oxidized) to primary brain cultures or to brain in vivo in the absence or presence of either apoE epsilon 2, epsilon 3, or epsilon4 isoforms or apoJ (which, in turn, will be supplied either exogenously or via transgenesis). In Project 2, levels of circulating soluble obtained from normal subjects, elderly individuals and AD patients. sAbeta distribution among lipoprotein particles and the influence of apoE isoforms on sAbeta distribution will also be determined. Both non-oxidized and oxidized apolipoproteins will be studied. In relate aims, the cellular sites of clearance and degradation of circulating sAbeta Project 3, models of cerebral amyloid angiopathy will be created using novel as well as proven strategies. Also, constructing transgenic mice over-expressing the human betaAPP gene in neuronal cells, using the human betaAPP cDNA, containing the FAD K670N/M671L mutations, driven by murine Thy-1 promoter or the ARPP-21 neuronal specific core promoter. Finally, transgenic mice over-expressing the human apoL protein in glial cells using the glial fibrillary acidic protein promoter will be produced and used to determine the role of apoJ in the solubilization of Abeta. These Projects will be supported by an Administrative Core, a Histopathology Core, and an Abeta Core. The "Abeta Core" will oversee standardization of Abeta structural states and specialized, highly sensitive measurements of Abeta levels.