Proteomics/Neuropathology Core

蛋白质组学/神经病理学核心

• 批准号: 10428581
• 负责人: Beatrix Magdalena Ueberheide
• 金额: $ 25.54万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10428581
• 关键词: 3xTg-AD mouse; AD transgenic mice; APP-PS1; Affect; Affinity Chromatography; Aftercare; Alzheimer' s Disease; Alzheimer' s disease pathology; Amyloid; Amyloid beta-Protein; Animal Disease Models; Antibodies; Antibody Therapy; Apolipoprotein E; Autopsy; Binding Proteins; Biology; Blood - brain barrier anatomy; Blood Vessels; Brain; Cerebral Amyloid Angiopathy; Cognitive; Collaborations; Data; Data Set; Development; Drug usage; Ensure; Equipment; Freezing; Goals; Health; Human; Human Resources; Immunoglobulin G; Immunoglobulin M; Immunotherapy; Injections; Label; Laboratories; Lesion; Mass Spectrum Analysis; Molecular; Monoclonal Antibodies; Mus; Neurodegenerative Disorders; Neurologist; Neurosciences; Passive Immunotherapy; Pathogenesis; Pathway interactions; Patients; Peptides; Peptoids; Pharmaceutical Preparations; Protein Analysis; Protein Isoforms; Proteins; Proteome; Proteomics; Publications; Reproducibility; Resources; Role; Sampling; Senile Plaques; Standardization; Technical Expertise; Therapeutic; Therapeutic antibodies; Tissue Sample; Tissues; Transgenic Mice; Work; apolipoprotein E-2; apolipoprotein E-3; apolipoprotein E-4; behavioral study; beta pleated sheet; brain tissue; clinical practice; data integration; experience; human tissue; instrumentation; laser capture microdissection; mild cognitive impairment; mouse model; neuropathology; novel; novel strategies; pre-clinical; small molecule; tau-1

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.

核心B-摘要/摘要 阿尔茨海默氏病（AD）病理是异质的，但对不同的了解 缺少驱动AD病理的途径。核B的作用是提供公正的表征 在AD发病机理和治疗方法中受不同APOE同工型影响的蛋白质和途径 针对这个角色。核心B将提供来自纽约大学大脑库的人类验尸组织的表现良好 并从阿尔茨海默氏病中心（RADC）收到验尸组织。核心将提供 神经病变病变蛋白质组的无偏表征（实质和血管淀粉样蛋白） 使用我们最近建立的局部蛋白质组学方法 - 激光捕获微分解（LCM） 然后进行无标签定量质谱法（LC-MS）。样品由从中收集的组织组成 APOE4，APOE3或APOE2载体的AD患者具有全谱AD（临床前） 认知正常，轻度认知障碍（MCI）和AD）以及表达的AD转基因小鼠模型 人类APOE 2,3或4个同工型。此外，核心将验证项目2中使用的肽和小药物 通过注入小鼠之前的质谱法。该核心将提供一个最先进的分析质量 光谱平台将用于3个项目。此关键功能将确保获得 数据是可重现的，并促进了将所有3个项目的发现相关联的下游分析。这 核心的具体目的是： 1。提供使用标准化的最新神经病理学来表征的人类脑组织 分析（项目1和3） 2。表征和量化淀粉样斑块和脑淀粉样血管病（CAA） 临床前认知正常，MCI和APOE4，APOE3和APOE2载体的蛋白质组织 （Proje CT 1）。 3。使用亲和力纯化来表征Aβ和磷酸化的tau结合蛋白 由MS（项目1）。 4。表征表达APOE2的转基因小鼠中的淀粉样菌斑和CAA蛋白 APOE3，APOE4或APOE KO，有无处理（肽，小分子药物） （Proje CT 2）。 5。验证项目3中开发的治疗抗体是否越过血脑屏障 （Proje CT 3）。 6。表征IgM和IgGAβComab对CAA的被动免疫疗法的影响 tgswdi，3xtg，app/ps1小鼠的蛋白质组越过APOE2，E3和E4背景 （Proje CT 3）。 核心B产生的蛋白质组学数据将有助于对APOE在 AD的发病机理，将有助于发现与AD开发有关的蛋白质和途径。 重要的是，人体组织和AD动物模型的联合使用将增强 临床实践的发现。