Structure of amyloid fibrils in human neurodegenerative diseases and aging

项目总结/摘要 最近，在了解tau蛋白病以及tau蛋白聚集体如何导致神经退行性变方面取得了重大进展， 通过描述人脑中tau纤维的原子结构，我们有 确定了疾病中包含所有六种tau亚型（3R+4 R tau）的tau细丝的形态 与细胞外淀粉样蛋白沉积相关的蛋白质是相同的，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起tau应答，包括 3R+4 R tau蛋白，具有一个共同的折叠（AD折叠），我们表征了人细胞外 阿尔茨海默病（AD）中的淀粉样蛋白-β（Aβ）和朊病毒疾病中的朊病毒蛋白淀粉样蛋白（APrP）。另外我们 表征了弥漫性路易体病、帕金森病和帕金森病中α-突触核蛋白丝的结构， 多系统萎缩和TMEM 106 B细丝在几种神经退行性疾病和衰老中的作用。的 在这个MPI应用程序中提出的研究是这一开创性工作的逻辑延续。这些 研究是对PAR-22-208“阿尔茨海默病相关痴呆（ADRDs）的结构生物学”的回应。 蛋白质病”（R 01） 本申请的主要目标是继续从脑组织中表征脑纤维的结构， 高原子级分辨率，用于未来PET配体开发和其他相关用途， 强调突变/基因型和散发性与家族性疾病，不同的发病年龄（例如， 早发与迟发），以及共存病变的存在。此外，我们建议表征纤维来源于 体内来源如相关的动物模型和生物学相关的体外系统，并合成， 鉴定和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的 目标。第一个是对具有不同APOE的EOAD个体的大型队列进行冷冻EM研究 基因型和发病年龄，以及具有不同临床、遗传和神经病理特征的家族性EOAD个体。 功能.此外，我们还将首次确定唐氏症个体的细丝结构 综合征，家族性英国和丹麦痴呆症和Biondi体的结构和身份。二是 生成冷冻EM图并确定从大脑中提取的细丝的相应原子模型 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cryo-EM，重组tau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。

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.