Role of ApoE4 Domain Interaction in Neurodegeneration

ApoE4 结构域相互作用在神经退行性变中的作用

• 批准号: 6835132
• 负责人: KARL WEISGRABER
• 金额: $ 39.29万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835132
• 关键词: apolipoprotein E; astrocytes; intermolecular interaction; laboratory mouse; neural degeneration; protein binding; protein structure function; structural biology; tissue /cell culture

DESCRIPTION (provided by the applicant): Apolipoprotein (apo) E4 is an established risk factor for neurodegenerative disease, including Alzheimer's disease (AD), and for poor outcome from head trauma and stroke. However, the mechanism underlying this increased risk remains elusive. Since protein function is directly related to protein structure, we have focused on determining the structural features that distinguish the apoE isoforms to gain insight into how these differences relate to the mechanism for the different effects of the isoforms in neurodegeneration. Our structural and mutagenesis studies established that apoE contains two structural domains. In apoE4, but not apoE3 and apoE2, the two domains interact. Our working hypothesis is that this unique structural property of apoE4 has a major influence on its functional properties, including lipid transport, metabolism, and mechanisms by which apoE4 contributes to neurodegeneration and heart disease. The aims in this application are designed to test this hypothesis in the context of neurodegeneration and AD using in vitro model systems and a novel apoE mouse model, in which domain interaction was engineered into mouse apoE in by gene targeting (Arg-6 1 mouse apoE). In Specific Aim 1, we will test the hypothesis that domain interaction determines the lipid-binding properties of Arg-6 1 mouse apoE and human apoE4. In Specific Aim 2, we will test the hypothesis that domain interaction in Arg-61 mouse apoE influences the type and composition of lipoprotein particles secreted by cultured primary astrocytes. In Specific Aim 3, we will test the hypothesis that domain interaction in Arg-6 1 mouse apoE decreases neuronal outgrowth in cell and organ culture systems. In Specific Aim 4, we will test the hypothesis that domain interaction in Arg-6 1 mouse apoE4 contributes to neurodegeneration. The results from these studies have the potential to provide clues into the mechanism by which apoE4 contributes to neurodegeneration and to identify therapeutic targets based on isoform-specific effects.

描述（申请人提供）：apoleipoylotin（apo）E4是 神经退行性疾病的确定危险因素，包括阿尔茨海默氏症 疾病（AD），以及头部外伤和中风的不良结果。但是， 这种增加的风险基础的机制仍然难以捉摸。由于蛋白质 功能与蛋白质结构直接相关，我们专注于 确定区分APOE同工型获得的结构特征 深入了解这些差异如何与不同的机制相关联 同工型在神经变性中的影响。我们的结构和诱变 研究确定APOE包含两个结构域。在apoe4中，但是 不是APOE3和APOE2，两个域相互作用。我们工作的假设是 APOE4的独特结构特性对其具有重大影响 功能特性，包括脂质转运，代谢和机制 APOE4有助于神经变性和心脏病。目标 该应用程序旨在在以下情况下检验该假设 使用体外模型系统和新型ApoE小鼠的神经变性和AD 模型，其中域相互作用被基因设计为小鼠APOE 靶向（ARG-6 1鼠标APOE）。在特定目标1中，我们将检验假设 该域相互作用决定了arg-6 1的脂质结合特性 鼠标apoe和人类apoe4。在特定目标2中，我们将检验以下假设。 arg-61小鼠APOE中的域相互作用会影响 由培养的原代星形胶质细胞分泌的脂蛋白颗粒。在特定目标中 3，我们将检验以下假设：arg-6 1小鼠apoE中的域相互作用 降低细胞和器官培养系统中的神经元出生。在特定目标中 4，我们将检验以下假设：arg-6 1小鼠apoe4中的域相互作用 有助于神经变性。这些研究的结果具有 有可能向APOE4贡献的机制提供线索的潜力 神经变性并确定基于同工型特异性的治疗靶标 效果。