Autophagy and Neurodegeneration

描述（由申请人提供）：项目概述在过去的几年中，巨细胞自噬已经与从聚集倾向性疾病亨廷顿病到溶酶体贮积症Neiman-Pick C型的广泛的神经退行性疾病有关。然而，尽管它的流行，宏自噬仍然知之甚少，使得很难定义它如何有助于发病机制。也许不足为奇的是，在不同的疾病中，巨自噬被认为是疾病进展中的潜在致病因素和潜在改善因素。如果我们要针对这种复杂的降解途径进行治疗，我们需要更好地定义自噬过程，以便将其应用于大脑。在这次拨款申请中，我们建议通过关注所涉及的关键细胞器：自噬泡（AV）来获得对宏观自噬的新见解。被定义为一个洋葱样的多层囊泡，对标志物MAP 1 LC 3（ATG 8的哺乳动物同源物）呈阳性，这种结构的形成和成熟是自噬过程的核心，迄今为止是最不了解的。使用我们已经开发的可以分离特定AV群体用于蛋白质组学和基于脂质的分析的新方法，我们将：1）表征来自神经元细胞和脑的AV; 2）比较和对比来自用其他四种ATG 8哺乳动物同源物标记的囊泡的MAP 1 LC 3标记的AV;和3）使用基于功能细胞的测定来进一步确定各种ATG 8蛋白质组如何影响大自噬。公共卫生相关性：大自噬是一个知之甚少的过程，它对于允许细胞（如神经元）摆脱不再起作用的蛋白质非常重要。有趣的是，这一过程被认为是许多神经退行性疾病的核心，例如亨廷顿病、帕金森病、阿尔茨海默病、许多溶酶体贮积病等。在这里，我们建议研究与大脑相关的宏观自噬，以便我们可以使用这些信息来设计针对这些疾病的有效治疗方法。

DESCRIPTION (provided by applicant): Project Summary Over the last several years, macroautophagy has been implicated in a wide array of neurodegenerative disorders from the aggregation prone disorder, Huntington's disease to the lysosomal storage disorders, Neiman-Pick Type C. Despite its prevalence however, macroautophagy is still poorly understood, making it difficult to define how it contributes towards pathogenesis. Perhaps unsurprisingly, in different disorders, macroautophagy has been considered both as a potentially causative and potentially ameliorative element in disease progression. If we are to target this complex degradative pathway for therapeutics, we need to better define the autophagic process in a means we can apply it towards the brain. In this grant submission, we propose to gain new insights into macroautophagy by focusing on the key organelle involved: the autophagic vacuole (AV). Defined as an onion-like multilamellar vesicle that is positive for the marker MAP1LC3 (a mammalian homologue of ATG8), the formation and maturation of this structure is at the heart of the autophagic process and is by far the least understood. Using a novel approach which we have developed that can isolate specific populations of AV for proteomic and lipid-based analyses, we will: 1) characterize AVs from neuronal cells and brain; 2) compare and contrast MAP1LC3- labeled AVs from vesicles labeled with the other four ATG8 mammalian homologues; and 3) use functional cell based assays to further define how the various ATG8- proteomes impact macroautophagy. PUBLIC HEALTH RELEVANCE: Macroautophagy is a poorly understood process that is important for allowing cells, such as neurons to get rid of proteins that no longer function. Interestingly, this process has been implicated to be at the heart of many neurodegenerative diseases such as Huntington's disease, Parkinson's disease, Alzheimer's disease, many lysosomal storage diseases and others. Here we propose to study macroautophagy as it pertains to the brain so that we can use this information to design effective treatment for these many diseases.