Proteomics/Neuropathology Core

核心B--摘要/摘要 阿尔茨海默病(AD)的病理是异质性的，但对不同的 推动AD病理的通路缺失。核心B的作用是提供对 不同载脂蛋白E亚型对AD发病机制及治疗途径的影响 瞄准这个角色。核心B将提供来自纽约大学脑库的具有良好特征的人类尸检组织 以及从拉什阿尔茨海默病中心(RADC)收到的尸检组织。核心B将提供 神经病理损伤(实质和血管淀粉样蛋白)蛋白质组的无偏特征 使用我们最近建立的定位蛋白质组学方法-激光捕获显微切割(LCM) 然后是无标记定量质谱仪(LC-MS)。这些样本由收集自 具有全谱AD(临床前)的apoE4、apoE3或apoE2携带者的AD患者 认知正常、轻度认知障碍(MCI)和AD)以及表达表达的AD转基因小鼠模型 人类载脂蛋白E 2、3或4亚型。此外，核心将核实项目2中使用的类肽类药物和小药物 在注射到小鼠体内之前通过质谱分析。这个核心将提供单一的最先进的分析质量 光谱平台，将在所有3个项目使用。这一关键功能将确保收购的 数据是可重现的，便于对所有3个项目的结果进行相互关联的下游分析。这个 核心的具体目标是： 1.提供以标准化最先进的神经病理学为特征的人脑组织 分析(项目1和3) 2.淀粉样斑块和脑淀粉样血管病(CAA)的特征和定量 ApoE4、apoE3和apo2携带者的临床前认知正常、MCI和晚期AD的蛋白质组 (项目ct 1)。 3.用亲和纯化方法鉴定Aβ和磷酸化tau结合蛋白 MS(项目1)。 4.鉴定表达apoE2转基因小鼠的淀粉样斑块和CAA蛋白质组。 ApoE3、apoE4或apoE KO，接受和不接受治疗(类肽、小分子药物) (项目ct 2)。 5.验证项目3中开发的治疗性抗体是否能穿过血脑屏障 (项目ct 3)。 6.β联合抗体被动免疫治疗对慢性再生障碍性贫血的影响 TgSwDI、3xTg、APP/PS1小鼠的蛋白质组交叉到APOE2、E3和E4背景上 (项目ct 3)。 核心B产生的蛋白质组数据将有助于更好地理解载脂蛋白E在 并将有助于发现参与阿尔茨海默病发展的蛋白质和途径。 重要的是，人类组织和AD动物模型的结合使用将增强 将发现转化为临床实践。

CORE B- SUMMARY/ABSTRACT Alzheimer’s Disease (AD) pathology is heterogenous, yet a comprehensive understanding of the different pathways that drive AD pathology is missing. The role of core B is to provide an unbiased characterization of proteins and pathways affected by different apoE isoforms in AD pathogenesis and therapeutic approaches targeting this role. Core B will provide well characterized human post mortem tissue from the NYU brain bank and post mortem tissue received from the Rush Alzheimer’s Disease Center (RADC). Core B will provide the unbiased characterization of the proteome of neuropathological lesions (parenchymal and vascular amyloid) using our recently established approach of localized proteomics - Laser Capture Microdissection (LCM) followed by label-free quantitative mass spectrometry (LC-MS). The samples consist of tissues collected from patients with AD who are apoE4, apoE3 or apoE2 carriers that have the full spectrum of AD (preclinical cognitive normal, mild cognitive impairment (MCI) and AD), and AD transgenic mice models that express human ApoE 2,3 or 4 isoforms. In addition, the core will verify the peptoids and small drugs used in Project 2 by mass spectrometry prior to injection into mice. This core will provide a single state of the art analytical mass spectrometry platform that will be used across all for 3 projects. This key feature will ensure that the acquired data is reproducible and facilitates the downstream analysis of correlating the findings of all 3 projects. The specific aims of the Core are: 1. Provide human brain tissue characterized using standardized state-of-the-art neuropathological analysis (Projects 1 and 3) 2. Characterize and quantify the amyloid plaque and cerebral amyloid angiopathy (CAA) proteomes of preclinical cognitive normal, MCI, and late AD of apoE4, apoE3 and apoE2 carriers (Proje ct 1). 3. Characterize Aβ and phosphorylated tau binding proteins using affinity purifications followed by MS (Project 1). 4. Characterize the amyloid plaque and CAA proteomes in transgenic mice expressing apoE2, apoE3, apoE4 or apoE KO, with and without treatment (peptoid, small molecule drugs) (Proje ct 2). 5. Verify that the therapeutic antibodies developed in Project 3 cross the blood brain barrier (Proje ct 3). 6. Characterize the effect of passive immunotherapy with IgM and IgG AβComAbs on the CAA proteome in TgSwDI, 3xTg, APP/PS1 mice crossed onto an apoE2, E3 and E4 background (Proje ct 3). The proteomic data generated by Core B will facilitate greater understanding of apoE’s role in the pathogenesis of AD and will aid the discovery of proteins and pathways involved in the development of AD. Importantly, the combined use of human tissue and AD animal models will enhance the translatability of the findings to clinical practice.