Integrated Platform to study Neurodegeneration in Alzheimer’s Disease

项目总结/摘要 在阿尔茨海默氏病（AD）中，细胞内后修饰的Tau聚集体的积累是阿尔茨海默氏病（AD）的一个重要特征。 AD病理学的标志和认知损害的强相关性。当十几个 病理性聚集体中Tau蛋白的翻译后修饰（PTM）已经研究了几十年，我们的研究发现， 新的定量和定性质谱方法已经在角型中发现了~100个PTM。 来自98名人类阿尔茨海默病患者的脑回在这些聚集体中，即Braak阶段V/VI，和年龄 对照组额叶无病理改变。在这份提案中，我们的目标是发展 一个大规模的蛋白质组学平台，用于绘制疾病进展时PTM的时间发生。我们将 检查一组有价值的新病例，选择涵盖从0到VI的所有Braak阶段。所有案件在此 新的队列将有详细的死前临床数据，这将使我们能够检测和评估相关性 临床和病理生理学发现与本研究中鉴定的任何tau PTM之间的差异。为此 分析，我们将研究3个脑区（内嗅/杏仁核，颞叶和视觉皮层）在20 Braak 0 受试者; 40例早期Braak I/II期受试者; 40例中期Braak III/IV期受试者; 40例晚期Braak V/VI期受试者 科目我们还将绘制这名特征明确的患者大脑标本的整体蛋白质组 以鉴定与这些Braak阶段依赖性PTM相关的酶。Tau的发现 将使用广泛接受的Tau传感器测定来验证蛋白质组的表征和作图 用于研究AD中的Tau聚集。 总之，该Tau PTM特异性数据集、深度蛋白质组图谱和后续验证 播种试验中的实验将用于测试我们的总体假设：了解 Tau PTM的进行性积累导致Tau的毒性和播种能力，并绘制Tau的基因图谱。 这些PTM和负责它们的酶的疾病进展依赖性将使我们能够开发 在疾病的早期和前驱阶段的干预策略。

Project Summary/Abstract In Alzheimer's disease (AD), the accumulation of intracellular posttranslationally modified Tau aggregates, is a hallmark of AD pathology and a strong correlate of cognitive impairment. While the dozen or so Posttranslational Modifications (PTMs) on Tau in pathological aggregates have been studied for decades, our new quantitative and qualitative mass spectrometry approaches have discovered ~100 PTMs in the angular gyrus from 98 human Alzheimer's Disease patients in these aggregates, i.e. Braak stages V/VI, and age matched control subjects without pathological changes in the frontal lobe. In this proposal, we aim to develop a large-scale proteomics platform to map the temporal occurrence of PTMs as disease progresses. We will examine a valuable new set of cases, selected to encompass all Braak stages from 0 to VI. All cases in this new cohort will have detailed premortem clinical data, which will allow us to detect and assess correlations between clinical and pathophysiological findings and any tau PTM that is identified in this study. For this analysis, we will study 3 brain regions (entorhinal/amygdala, and temporal and visual cortex) in 20 Braak 0 subjects; 40 early stage Braak I/II subjects; 40 mid stage Braak III/IV subjects; and 40 end stage Braak V/VI subjects. We will also map the global proteomes of the brain specimens from this well-characterized patient cohort to identify the enzymes associated with these Braak stage dependent PTMs. The findings of the tau characterization and mapping of the proteomes will be validated using widely accepted Tau sensor assays used to study Tau aggregation in AD. In summary, this Tau PTM specific dataset, the deep proteome maps and the subsequent validation experiments in seeding assays will be used to test our overarching hypothesis: Understanding how the progressive accumulation of Tau PTMs results in the toxicity and seeding competence of tau, and mapping the disease progression dependence of these PTMs and the enzymes responsible for them will allow us to develop interventional strategies at earlier and prodromal stages of disease.