A quantitative framework for understanding endosomal trafficking networks in Alzheimer's disease

了解阿尔茨海默氏病内体运输网络的定量框架

基本信息

批准号：
10470286
负责人：
JEFFREY W HARPER
金额：
$ 45.94万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10470286
关键词：
Abeta clearance Abeta synthesis Affect Alzheimer&apos s Disease Amyloid beta-Protein Architecture Autophagocytosis Biochemical Biological Biological Models Biotinylation CRISPR screen Candidate Disease Gene Cell Line Cell membrane Cells Defect Degradation Pathway Dependence Disease Event Excision Genes Genetic Goals Golgi Apparatus Image Individual Laboratories Link Lysosomes Maps Membrane Proteins Methodology Mitochondria Molecular Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Organelles PINK1 gene Pathogenesis Pathway Analysis Pathway interactions Predisposition Prevention Process Proteins Proteomics Quality Control Recycling Reporter Role Sorting - Cell Movement Specificity System Testing Therapeutic Time Toxic effect Ubiquitin Validation Work abeta accumulation abeta toxicity arm base cell type design experimental study gain of function mutation gene network genetic analysis human embryonic stem cell insight late endosome misfolded protein multicatalytic endopeptidase complex mutant neuropathology novel protein degradation protein misfolding protein transport proteostasis repaired screening tissue/cell culture trafficking

项目摘要

PROJECT SUMMARY Accumulating evidence suggest that neurodegenerative diseases including Alzheimer’s disease (AD) are frequently the result of alterations in endosomal trafficking and proteostasis pathways, one consequence of which can be the accumulation of aggregation-prone proteins. Numerous familial and sporadic loss or gain-of- function mutations have been identified in such pathways, illuminating potential drivers of disease pathogenesis. In addition, excess protein mis-folding due to altered trafficking could affect proteostasis pathways by blocking protein turnover or trafficking. Protein and organelle trafficking within cells is a highly dynamic and interconnected process, and defects in one arm of the system can affect other aspects of the network in unpredictable ways including reduced flux in turnover pathways. Indeed, it is conceivable that many seemingly unrelated mutations across the trafficking landscape in various neurodegenerative diseases reveal common mechanistic vulnerabilities downstream but with distinct cell-type sensitivities reflective of the identity of mis-trafficked proteins. As such, understanding the global architecture of trafficking systems and the key machinery that controls the directionality and efficiency of trafficking, particularly of aggregation-prone neurodegenerative proteins such as APP and its aggregation-prone form A, represents a central goal of the field. A aggregation as a toxic driver of AD neuropathology has been a dominant hypothesis in the field. However, thus far therapeutics directed at aggregate prevention or removal have not been successful, and alternative hypotheses including alterations in intracellular trafficking as an important event in neuropathology have emerged. Here, we seek to combine powerful genetic and proteomic approaches to develop a quantitative framework for understanding how disruption of major endosomal trafficking systems – retromer and retriever, found defective in neurodegenerative diseases – alter global membrane protein trafficking, and specific trafficking and processing of APP proteoforms. These studies make use of an extensive tool-kit of mutant tissue culture cell lines and induced neurons derived from human embryonic stem cell (hESC), in combination with targeted and unbiased proteomics of individual organelles linked with endosomal trafficking, to assemble a global map of cargo and trafficking dependencies. In parallel, we will employ novel flux-based screening strategies to search for genes controlling APP/A trafficking to the lysosome and the plasma membrane, and will examine the extent to which A accumulation within the endo-lysosomal system alters selective autophagic flux using new cargo-specific reporters. The central hypothesis being tested is that specific defects in protein trafficking networks underlies the susceptibility of neurons to A and other aggregation prone proteins and that these defects can be molecularly unmasked through systematic network and genetic analysis.

项目摘要积累的证据表明，但是包括阿尔茨海默氏病（AD）在内的神经发病障碍疾病是经常是内体贩运和蛋白质途径改变的结果，这是这可能是易于聚集的蛋白的积累。在这种途径中已经确定了功能，从而照亮了疾病的潜力发病机理。此外，由于运输的改变而过多的蛋白质折叠可能会影响蛋白质通过阻止蛋白质周转或运输的途径。动态和国际过程以及系统的一个部门的缺陷可能会影响网络以不可预测的方式包括在流动途径中减少通量。在各种神经退行性疾病中，整个贩运景观中看似无关的叛变揭示了下游的常见机械脆弱性，但具有不同的细胞类型敏感性反映了身份贩运蛋白质的错误。控制贩运的方向性和效率的机械，特别是容易发生的机械神经退行性蛋白（例如App及其易于聚集）形成A，压抑了其中的中心。 AD。但是，到目前为止，用于预防或删除的治疗剂尚未成功，并且替代假设，包括改变细胞内贩运的改变作为神经囊肿的重要事件出现了。定量框架，用于了解主要内体贩运系统如何破坏 - 逆转录器和猎犬发现神经退行性疾病有缺陷 - 改变全球膜蛋白运输，和这些研究的特定运输和处理。突变组织培养细胞lins和诱导源自人类胚胎干细胞（HESC）的神经元，与内体贩运相关的个体细胞器的靶向和公正蛋白质组学结合，为了组装全球货物和贩运依赖地图。筛选搜索控制App/A运输溶酶体和等离子体的基因的策略膜，并将检查在内部溶酶体系统中A积累的程度选择性的自噬通量使用新的货物特异性记者。蛋白质运输网络中的特定缺陷是神经元对A和其他的敏感性聚集易于蛋白质，这些缺陷缺陷可以是分子的和遗传分析。