Proteomics/Neuropathology Core

蛋白质组学/神经病理学核心

基本信息

批准号：
10621829
负责人：
Beatrix Magdalena Ueberheide
金额：
$ 27.74万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
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/10621829
关键词：
3xTg-AD mouse AD transgenic mice APP-PS1 Affect Affinity Chromatography Aftercare Alzheimer&apos s Disease Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Amyloid Amyloid beta-Protein Animal Disease Models Antibodies Apolipoprotein E Autopsy Binding Proteins Biology Blood Vessels Board Certification 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 blood-brain barrier crossing 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

项目摘要

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/摘要阿尔茨海默氏病 (AD) 的病理学具有异质性，但对不同病理学的全面了解驱动 AD 病理学的途径缺失核心 B 的作用是提供公正的表征。 AD发病机制和治疗方法中受不同apoE亚型影响的蛋白质和通路 Core B 将提供来自纽约大学脑库的特征明确的人类死后组织。从 Rush 阿尔茨海默病中心 (RADC) 收到的死后组织将提供该信息。神经病理病变蛋白质组的公正表征（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 - 激光捕获显微切割 (LCM) 随后进行无标记定量质谱分析 (LC-MS)。样品由采集自的组织组成。患有全谱 AD 的 apoE4、apoE3 或 apoE2 携带者的 AD 患者（临床前认知正常、轻度认知障碍 (MCI) 和 AD）以及 AD 转基因小鼠模型人类ApoE 2,3或4亚型此外，核心还将验证项目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分析该核心将提供单一的最先进的分析质量。将在所有 3 个项目中使用的光谱测量平台将确保所获得的。数据是可重复的，有助于下游分析所有 3 个项目的结果。核心的具体目标是： 1. 提供使用标准化的最先进的神经病理学表征的人类脑组织分析（项目1和3） 2. 淀粉样斑块和脑淀粉样血管病 (CAA) 的表征和定量 apoE4、apoE3 和 apoE2 携带者临床前认知正常、MCI 和晚期 AD 的蛋白质组（项目1）。 3. 随后使用亲和纯化表征 Aβ 和磷酸化 tau 结合蛋白由 MS（项目 1）提供。 4. 表达 apoE2 的转基因小鼠中淀粉样斑块和 CAA 蛋白质组的特征， apoE3、apoE4 或 apoE KO，有或没有治疗（类肽、小分子药物）（项目2）。 5.验证项目3开发的治疗性抗体是否能穿过血脑屏障（项目3）。 6. 表征 IgM 和 IgG AβComAb 被动免疫疗法对 CAA 的影响 TgSwDI、3xTg、APP/PS1 小鼠中的蛋白质组交叉到 apoE2、E3 和 E4 背景上（项目3）。 Core B 生成的蛋白质组数据将有助于更好地了解 apoE 在 AD 的发病机制，并将有助于发现与 AD 发展有关的蛋白质和途径。重要的是，人体组织和 AD 动物模型的结合使用将增强该研究的可翻译性。研究结果应用于临床实践。