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-总结/摘要 阿尔茨海默病（AD）病理学是异质的，但对不同病理学的全面理解是不一致的。 驱动AD病理学的途径缺失。核心B的作用是提供一个公正的表征， 在AD发病机制和治疗方法中受不同apoE亚型影响的蛋白质和途径 瞄准这个角色。核心B将提供来自纽约大学脑库的充分表征的人类死后组织 以及从拉什阿尔茨海默病中心（RADC）获得的死后组织。核心B将提供 神经病理学病变（实质和血管淀粉样蛋白）蛋白质组的无偏表征 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） 随后进行无标记定量质谱法（LC-MS）。样本由从以下组织中收集的组织组成： 具有AD全谱（临床前）的apoE 4、apoE 3或apoE 2携带者的AD患者 认知正常、轻度认知障碍（MCI）和AD），以及表达 人Apoe 2、3或4亚型。此外，核心将验证项目2中使用的类肽和小药物 在注射到小鼠体内之前通过质谱法测定。这个核心将提供一个单一的最先进的分析质量 光谱分析平台，将用于所有3个项目。这一关键特征将确保所收购的 数据是可重复的，便于下游分析，将所有3个项目的调查结果联系起来。的 核心的具体目标是： 1.提供使用标准化最先进的神经病理学表征的人脑组织 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） apoE 4、apoE 3和apoE 2携带者的临床前认知正常、MCI和晚期AD的蛋白质组 （项目1）。 3.使用亲和纯化表征Aβ和磷酸化tau结合蛋白， 作者：MS（项目1）。 4.表征表达apoE 2的转基因小鼠中的淀粉样斑块和CAA蛋白质组， apoE 3、apoE 4或apoE KO，伴或不伴治疗（类肽、小分子药物） （项目2）。 5.验证项目3中开发的治疗性抗体穿过血脑屏障 （项目3）。 6.用IgM和IgG AβComAbs对CAA进行被动免疫治疗的效果表征 在apoE 2、E3和E4背景上杂交的TgSwDI、3xTg、APP/PS1小鼠中的蛋白质组 （项目3）。 Core B产生的蛋白质组学数据将有助于更好地理解apoE在细胞凋亡中的作用。 本发明的目的在于揭示AD的发病机制，并将有助于发现参与AD发展的蛋白质和途径。 重要的是，人组织和AD动物模型的组合使用将增强本发明的可翻译性。 临床实践的发现。