AUTOPHAGY FUNCTION & DYSFUNCTION IN ALZHEIMER'S DISEASE

自噬功能

• 批准号: 6920487
• 负责人: RALPH A. NIXON
• 金额: $ 34.01万
• 依托单位: NATHAN S. KLINE INSTITUTE FOR PSYCH RES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6920487
• 关键词: Alzheimer' s disease; Downs syndrome; RNA interference; amyloid proteins; aspartic endopeptidases; autophagy; cell line; cellular pathology; endocytosis; enzyme activity; fibroblasts; gene deletion mutation; gene mutation; genetically modified animals; green fluorescent proteins; immunoelectron microscopy; immunofluorescence technique; laboratory mouse; lysosomes; molecular pathology; neurons; neuropathology; neuroprotectants; presenilin; protein degradation

Autophagy is the cell's primary mechanism to degrade damaged organelles and cytoplasm and to protect itself from toxic protein aggregates not degradable by the proteasome. We will address new leads revealing induction and impairment of autophagy in Alzheimer's Disease (AD), identifying its role in Abeta generation, and showing for a requirement for presenilin (PS1) in autophagy function. The hypothesis that autophagy is impaired in AD will be investigated in human brain and FAD fibroblasts, cell lines, and mouse models of AD pathology (Aims 1 and 2). Novel antibodies marking specific compartments of the autophagic pathway and other organelles will be used in double-immunofluorescence and ultrastructural studies (including immunogold), along with other state-of-the-art techniques, to define the nature and significance of autophagy dysfunction in AD. The roles of PS1, PS2, and g-secretase activity in modulating autophagy will be determined and the basis for the observed selective impairment in autophagy-mediated protein degradation caused by FAD-related PS1 mutations will be investigated (Aim 2). We will also investigate the basis for distinctive autophagy abnormalities in Trisomy 21 (Down syndrome) and test the hypothesis that FAD-related APP mutations (Aim 3) also impair autophagy. Autophagy as a neuroprotective mechanism will be studied in primary neurons from normal mice, AD mouse models, and a novel transgenic mouse expressing GFP-tagged LC3, a protein associated with autophagy induction, which will enable real-time visualization of autophagy and will facilitate in vivo studies of autophagy modulation (Aim 4). As a prelude to modulating autophagy as a therapeutic intervention in neurodegenerative disease, we will investigate strategies to manipulate autophagy in the brain in vivo (Aim 4). These ongoing studies, the first to investigate autophagy systematically in AD, will provide a comprehensive assessment of the impact of autophagy and autophagy dysfunction and identify novel approaches to treat proteinopathies, including AD.

自噬是细胞降解受损细胞器和细胞质并保护自身免受蛋白酶体不能降解的有毒蛋白质聚集体的主要机制。我们将讨论揭示阿尔茨海默病（AD）中自噬诱导和损伤的新线索，确定其在Abeta生成中的作用，并显示自噬功能中对早老素（PS1）的需求。将在人脑和FAD成纤维细胞、细胞系和AD病理学小鼠模型中研究AD中自噬受损的假设（目的1和2）。标记自噬途径和其他细胞器的特定隔室的新型抗体将用于双重免疫荧光和超微结构研究（包括免疫金），沿着其他最先进的技术，以确定自噬途径的性质和意义。 自噬功能障碍将确定PS1、PS2和g-分泌酶活性在调节自噬中的作用，并研究FAD相关PS1突变引起的自噬介导的蛋白质降解中观察到的选择性损伤的基础（目的2）。我们还将研究21三体（唐氏综合征）中独特的自噬异常的基础，并检验FAD相关APP突变（Aim 3）也损害自噬的假设。自噬作为一种神经保护机制将在来自正常小鼠、AD小鼠模型和表达GFP标记的LC 3的新型转基因小鼠的原代神经元中进行研究，所述GFP标记的LC 3是一种与自噬诱导相关的蛋白质，其将使得能够实时检测自噬。 自噬的可视化，并将促进自噬调节的体内研究（目的4）。作为调节自噬作为神经退行性疾病的治疗干预的前奏，我们将研究在体内操纵脑中自噬的策略（目的4）。这些正在进行的研究是第一个系统研究AD中自噬的研究，将全面评估自噬和自噬功能障碍的影响，并确定治疗蛋白质病（包括AD）的新方法。