Structure of amyloid fibrils in human neurodegenerative diseases and aging

人类神经退行性疾病和衰老中淀粉样原纤维的结构

• 批准号: 10721721
• 负责人: BERNARDINO Francesco GHETTI
• 金额: $ 392.43万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10721721
• 关键词: Address; Age; Age of Onset; Aging; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amino Acid Sequence; Amyloid; Amyloid Fibrils; Amyloid Proteins; Amyloid beta-Protein; Amyloid deposition; Amyloidosis; Animal Model; Binding; Brain; Brain region; British; Classification; Clinical; Cryoelectron Microscopy; Dementia; Deposition; Development; Diffuse; Diffuse Lewy Body Disease; Disease; Down Syndrome; Early Onset Alzheimer Disease; Early Onset Familial Alzheimer' s Disease; Environment; Ependymal Cell; Face; Filament; Future; Genetic; Genotype; Goals; Gossypium; Grain; Human; In Vitro; Individual; Inherited; International; Knowledge; Lead; Ligands; Maps; Mass Spectrum Analysis; Modeling; Molecular; Morphology; Multiple System Atrophy; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Parkinson Disease; Pathology; Peptides; Pick Disease of the Brain; Positron-Emission Tomography; Post-Translational Protein Processing; PrP; Prion Diseases; Property; Protein Isoforms; Proteins; Recombinant Proteins; Recombinants; Resolution; Sampling; Scanning Electron Microscopy; Source; Specificity; Stains; Structure; Structure of choroid plexus; System; Tauopathies; Testing; Three-Dimensional Image; Time; Transgenic Mice; Transmission Electron Microscopy; Western Blotting; Wool; Work; alpha synuclein; amyloid structure; apolipoprotein E-4; autosome; brain tissue; cell type; cohort; extracellular; human model; in vivo; in vivo imaging; mRNA Differential Displays; mouse model; multidisciplinary; nanometer resolution; neurodegenerative dementia; neuropathology; novel; presenilin-1; protein TDP-43; protein aggregation; protein purification; protein structure; response; structural biology; synthetic peptide; tau Proteins; tau aggregation

Project Summary/Abstract Recently, significant progress in understanding tauopathies and how tau aggregates lead to neurodegeneration has been made by the description of the atomic structures of tau filaments from human brain. We have established that the morphology of tau filaments comprising of all six tau isoforms (3R+4R tau) in diseases associated with extracellular amyloid deposition is identical, regardless of the amino acid sequence of the amyloid subunit. To gain further understanding on how different amyloids elicit a tau response comprising of 3R+4R tau with a common fold (AD fold), we characterized the structures of filaments of human extracellular amyloid-β (Aβ) in Alzheimer disease (AD) and Prion protein amyloid (APrP) in Prion diseases. In addition, we characterized the structures of α-synuclein filaments in Diffuse Lewy body disease, Parkinson disease and Multiple system atrophy, and TMEM106B filaments in several neurodegenerative diseases and aging. The studies proposed in this MPI application are a logical continuation of this groundbreaking work. These studies are in response to PAR-22-208 “Structural Biology of Alzheimer's Disease Related Dementias (ADRDs) Proteinopathies” (R01) The main goals of this application are to continue to characterize, from brain tissue, the structure of filaments at high atomic-level resolution for future use in PET ligand development and other related purposes, with special emphasis on mutations/genotypes and sporadic versus familial forms of disease, different ages of onset (e.g. early vs. late onset), and presence of co-pathologies. In addition, we propose to characterize fibrils derived from in vivo sources such as relevant animal models and biologically relevant in vitro systems, and to synthesize, identify, and characterize amyloid-specific ligands. In order to achieve our goals, our proposal has three specific aims. The first is to perform cryo-EM studies on a large cohort of EOAD individuals with different APOE genotypes and age at onset, and familial EOAD individuals with different clinical, genetic and neuropathologic features. In addition, we will determine for the first time the structure of filaments in individuals with Down syndrome, familial British and Danish dementia and the structure and identity of Biondi bodies. Second, we will generate a cryo-EM map and determine the corresponding atomic models of filaments extracted from the brain of murine models and generate high-resolution 3D images of the amyloid structures. Lastly, we will characterize by cryo-EM the structure of recombinant tau and synthetic peptides homologous to Aβ, ABri and ADan and compare our results with the peptides isolated from human and animal models. We will also synthesize, identify, and characterize amyloid-specific ligands.

项目概要/摘要 最近，在了解 tau 蛋白病以及 tau 蛋白聚集如何导致神经退行性疾病方面取得了重大进展 是通过描述人脑中 tau 蛋白丝的原子结构而得出的。我们有 确定疾病中 tau 丝的形态由所有六种 tau 亚型 (3R+4R tau) 组成 与细胞外淀粉样蛋白沉积相关的物质是相同的，无论其氨基酸序列如何 淀粉样蛋白亚基。为了进一步了解不同的淀粉样蛋白如何引发 tau 反应，包括 具有共同折叠（AD折叠）的3R+4R tau，我们表征了人类细胞外丝的结构 阿尔茨海默病（AD）中的淀粉样蛋白-β（Aβ）和朊病毒病中的朊病毒蛋白淀粉样蛋白（APrP）。此外，我们 表征了弥漫性路易体病、帕金森病和 α-突触核蛋白丝的结构 多系统萎缩，以及多种神经退行性疾病和衰老中的 TMEM106B 细丝。这 该 MPI 应用中提出的研究是这项开创性工作的逻辑延续。这些 研究针对 PAR-22-208“阿尔茨海默病相关痴呆 (ADRD) 的结构生物学” 蛋白质病”(R01) 该应用的主要目标是继续从脑组织中表征丝状结构 高原子级分辨率，用于未来 PET 配体开发和其他相关目的，具有特殊的 强调突变/基因型、散发性与家族性疾病形式、不同的发病年龄（例如，发病年龄） 早发与晚发），以及是否存在合并症。此外，我们建议表征源自 体内来源，例如相关动物模型和生物学相关的体外系统，并合成， 识别并表征淀粉样蛋白特异性配体。为了实现我们的目标，我们的建议有三个具体内容 目标。第一个是对一大群具有不同 APOE 的 EOAD 个体进行冷冻电镜研究 基因型和发病年龄，以及具有不同临床、遗传和神经病理学特征的家族性 EOAD 个体 特征。此外，我们将首次确定患有唐氏综合症的个体的纤维结构 综合征、英国和丹麦家族性痴呆以及比昂迪体的结构和特性。其次，我们将 生成冷冻电镜图并确定从大脑中提取的细丝的相应原子模型 小鼠模型并生成淀粉样蛋白结构的高分辨率 3D 图像。最后，我们将描述 通过冷冻电镜分析重组 tau 蛋白和与 Aβ、ABri 和 ADan 同源的合成肽的结构 将我们的结果与从人类和动物模型中分离的肽进行比较。我们还将综合、识别、 并表征淀粉样蛋白特异性配体。