Defining Molecular Signatures Underlying Lysosomal Dysfunction in Alzheimer’s Disease

溶酶体是膜结合的降解区室，其分解来自内吞的大分子， 吞噬和自噬途径，并发挥关键代谢和信号枢纽的作用。积累 有证据表明，在阿尔茨海默病和其他神经退行性疾病中， 正确履行职能。然而，由于缺乏研究体内细胞器的工具， 在阿尔茨海默病的进展过程中， 疾病，并且所提出的损伤的确切分子性质尚不清楚。我们理解 溶酶体在疾病中的参与是进一步有限的，因为溶酶体是罕见的，构成<3%的溶酶体。 cell. 在这里，我们寻求联合收割机结合强大的，最先进的方法，包括最近开发的快速溶酶体 分离（LysoIP）和无偏倚的蛋白质组学和代谢组学分析，以确定溶酶体是否以及如何 在阿尔茨海默病的鼠模型中的体内变化。我们建议将重点放在分离的溶酶体， 神经元和小胶质细胞，我们认为这对疾病的病理学至关重要。复倒数 神经元和小胶质细胞之间的相互作用对于调节大脑中最重要的方面是必不可少的。 功能，我们假设这两种细胞类型中内溶酶体系统的改变 危及中枢神经系统的完整性在这里，在目的I中，我们提出定义溶酶体 阿尔茨海默病进展过程中神经元和小胶质细胞的变化产生动态变化 这些细胞中溶酶体蛋白和代谢物的图谱。这一目标将产生新的小鼠模型和鲁棒性。 能够从神经元和小胶质细胞快速分离溶酶体的方案。在目标2中，我们将验证 从目标1中筛选出生物信息学候选者，为未来的机械解剖铺平了道路。 拟议的研究利用创新技术和概念来解决基本的分子 阿尔茨海默病的病理生物学方面，建立体内溶酶体变化的综合图谱 在神经元和小胶质细胞中。我们相信，这项工作将阐明溶酶体生物学的新方面， 并有可能改变我们对阿尔茨海默病机制基础的理解， 为治疗这种毁灭性疾病的未来发展提供信息。

Lysosomes are membrane-bound degradative compartments that break down macromolecules from endocytic, phagocytic and autophagic pathways, and serve the role of key metabolic and signaling hubs. Accumulating evidence suggests that in Alzheimer's disease and other neurodegenerative disorders lysosomes fail to correctly perform their functions. However, due to the paucity of tools to study organelles in vivo, so far there has been no systematic assessment of lysosomal alterations during progression of Alzheimer's disease, and the exact molecular nature of the proposed impairments is not known. Our understanding of the involvement of the lysosome in the disease is further limited because lysosomes are rare, constituting <3% of the cell. Here, we seek to combine powerful, state-of-the-art approaches including recently developed rapid lysosomal isolations (LysoIPs) and unbiased proteomic and metabolomic analyses to determine if and how lysosomes change in vivo in murine models of Alzheimer's disease. We propose to focus on lysosomes isolated from neurons and microglia which we expect to be critical to the pathology of the disease. Complex reciprocal interactions between neurons and microglia are essential for regulation of the most important aspects of brain function, and we hypothesize that alterations of the endolysosomal systems in these two cell types compromise the integrity of the central nervous system. Here, in Aim I we propose to define lysosomal alterations in neurons and microglia over a time course of Alzheimer's disease progression generating a dynamic atlas of lysosomal proteins and metabolites in these cells. This aim will generate novel mouse models and robust protocols enabling rapid lysosomal isolations from neurons and microglia. In Aim 2 we will validate bioinformatically filtered candidates from Aim 1, paving the way for future mechanistic dissections. The proposed research utilizes innovative technologies and concepts to address fundamental molecular aspects of pathobiology of Alzheimer's disease, building a comprehensive atlas of in vivo lysosomal changes in neurons and microglia. We believe that this work will shed light on novel aspects of lysosomal biology in the brain and has the potential to transform our understanding of the mechanistic basis of Alzheimer's disease, informing future developments in the treatment of this devastating disorder.