INTERDISCIPLINARY APPROACH TO ALZHEIMER DRUG DISCOVERY

阿尔茨海默病药物发现的跨学科方法

• 批准号: 6362209
• 负责人: SAMUEL E. GANDY
• 金额: $ 84.41万
• 依托单位: NATHAN S. KLINE INSTITUTE FOR PSYCH RES
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6362209
• 关键词: Alzheimer's disease; amyloid proteins; apolipoproteins

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.

该项目的总体目标是应用生物化学、细胞生物学和转基因等方法，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, gene； APOE, protein）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节β -淀粉样蛋白的形成。由于APOE - epsilon4相关的AD是最常见的AD形式，我们计划重点识别APOE依赖性（特别是epsilon4-异构体依赖性）和apoj依赖性现象，这些现象可能成为治疗靶点。在项目1中，研究人员将测量apoE-异构体/Abeta复合物的形成和Abeta纤维形成，在没有或存在非变性apoE （epsilon 2， epsilon3或epsilon4 [ROS]， apoJ等）的情况下，这些被认为有助于体内β -淀粉样蛋白的形成。这些现象的作用将在它以不同的物理状态之一（可溶性、聚集/纤维化、氧化）提供给原代脑培养物或在体内的大脑中，在没有或存在apoE、epsilon 2、epsilon 3或epsilon4异构体或apoJ（反过来，apoJ将通过外源或转基因提供）的情况下确定。在项目2中，从正常受试者、老年人和AD患者中获得循环可溶性水平。sAbeta在脂蛋白颗粒中的分布以及apoE亚型对sAbeta分布的影响也将被确定。将研究非氧化和氧化载脂蛋白。在相关的目标中，循环sAbeta项目3的清除和降解的细胞位点，脑淀粉样血管病的模型将使用新的和经过验证的策略创建。同时，利用人类betaAPP cDNA，构建在神经元细胞中过表达人betaAPP基因的转基因小鼠，该基因含有由小鼠Thy-1启动子或ARPP-21神经元特异性核心启动子驱动的FAD K670N/M671L突变。最后，利用胶质纤维酸性蛋白启动子在胶质细胞中产生过表达人apoL蛋白的转基因小鼠，并用于确定apoJ在Abeta增溶中的作用。这些项目将由行政核心、组织病理学核心和Abeta核心提供支持。“Abeta核心”将监督Abeta结构状态的标准化和专门的、高度敏感的Abeta水平测量。