Integrated Platform to study Neurodegeneration in Alzheimer’s Disease

项目摘要/摘要 在阿尔茨海默病(AD)中，细胞内翻译后修饰的Tau聚集体的积累是一种 阿尔茨海默病的病理特征与认知障碍有很强的相关性。而十几个人 翻译后修饰(PTM)对病理性集合体中Tau的研究已经有几十年了，我们的 新的定量和定性质谱学方法在角质谱中发现了约100个PTM 98例人类阿尔茨海默病患者的脑回，即Braak V/VI期和年龄 额叶无病变的匹配对照组受试者。在这项提案中，我们的目标是开发 一个大规模的蛋白质组学平台，用于绘制随着疾病进展而发生的PTMS的时间分布。我们会 研究一组有价值的新案例，选择涵盖从0到VI的所有Braak阶段。 新的队列将有详细的尸检临床数据，这将使我们能够检测和评估相关性 临床和病理生理学结果与本研究中确定的任何tau PTM之间的差异。为了这个 分析，我们将研究3个大脑区域(内嗅觉/杏仁核，以及颞叶和视觉皮质)在20Braak 0 受试者；40名早期Braak I/II受试者；40名中期Braak III/IV受试者；以及40名末期Braak V/VI受试者 研究对象。我们还将绘制这个特征良好的患者脑组织样本的全球蛋白质组图。 队列以确定与这些Braak分期依赖的PTM相关的酶。Tau的调查结果 将使用被广泛接受的Tau传感器分析来验证蛋白质组的特征和图谱 用于研究AD中的Tau聚集。 综上所述，该Tau PTM特定数据集、深层蛋白质组图谱和后续验证 播种试验中的实验将被用来检验我们的总体假设：理解 Tau PTMS的逐渐积累导致了tau的毒性和播种能力，并绘制了 这些PTM和负责它们的酶的疾病进展相关性将使我们能够发展 疾病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.