INTERDISCIPLINARY APPROACH TO ALZHEIMER DRUG DISCOVERY

该计划的总体目标是应用生化、细胞生物学和转基因方法，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)和载脂蛋白J(ApoJ)的重要神经生物学活性，并阐明这些蛋白如何调控β-淀粉样蛋白的发生。由于apoE epsilon 4相关的AD是最常见的AD形式，我们计划重点识别apoE依赖(特别是epsilon 4异构体依赖)和apoJ依赖的现象，这些现象可能成为治疗的靶点。在项目1中，研究人员将在没有或存在非变性apoE epsilon 2、epsilon 3或epsilon 4[ROS]、apoJ等的情况下，测量apoE-异构体/Abeta复合体的形成和Abeta纤维形成。它们被认为在体内有助于β-淀粉样蛋白的形成。这些现象的作用将在以各种物理状态(可溶、聚集/纤化、氧化)之一提供给原代脑培养细胞或在没有或存在apoE epsilon 2、epsilon 3或epsilon 4亚型或apoJ的情况下提供给活体脑后确定(反过来，apoJ将由外源或通过转基因提供)。在项目2中，从正常受试者、老年人和AD患者获得循环可溶性水平。还将确定sAβ在脂蛋白颗粒中的分布以及载脂蛋白E亚型对sAβ分布的影响。非氧化型和氧化型载脂蛋白都将被研究。在相关的AIMS中，循环sAbeta项目3的清除和降解的细胞位置，大脑淀粉样血管病的模型将使用新的和已证实的策略来创建。此外，利用含有FAD K670N/M671L突变的人βAPP基因，构建在神经细胞中过表达人βAPP基因的转基因小鼠，该突变由小鼠Thy-1启动子或ARPP-21神经元特异性核心启动子驱动。最后，利用胶质纤维酸性蛋白启动子在胶质细胞中过表达人APOL蛋白的转基因小鼠将被用来确定apoJ在Abeta增溶中的作用。这些项目将得到一个行政核心、一个组织病理学核心和一个Abeta核心的支持。“Abeta核心”将监督Abeta结构状态的标准化和Abeta水平的专门、高度敏感的测量。

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.