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LINKING TAU FILAMENT STRUCTURE TO PHENOTYPIC DIVERSITY IN HUMAN TAUOPATHIES

将 TAU 丝结构与人类 TAU 病表型多样性联系起来

基本信息

批准号：
8663324
负责人：
Martin Margittai
金额：
$ 30.09万
依托单位：
UNIVERSITY OF DENVER (COLORADO SEMINARY)
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8663324
关键词：
Adult Alzheimer&apos s Disease Amyloid Biological Biological Assay Brain Cells Data Deposition Disease Disease Progression Electrons Filament Fluorescence Genetic Polymorphism Goals Growth Human Knowledge Lead Link Measurement Measures Mediating Methodology Methods Mission Modeling Molecular Molecular Conformation Monoclonal Antibodies Mutation Neurodegenerative Disorders Outcome Pathogenesis Pathway interactions Phenotype Phosphorylation Pick Disease of the Brain Play Prions Process Production Progressive Supranuclear Palsy Property Protein Isoforms Public Health Publishing Reporter Research Role Route Seeds Spectrum Analysis Structure System Tauopathies Testing Work acrylodan base biophysical techniques conformer disease phenotype drug discovery experience inhibitor/antagonist innovation insight mutant neurotoxic novel novel strategies novel therapeutics polypeptide research study small molecule tau Proteins tau aggregation tau-1

项目摘要

DESCRIPTION (provided by applicant): There is a fundamental gap in understanding of how the misfolding of tau relates to the enormous phenotypic diversity observed in human tauopathies. Persistence of this gap constitutes an important problem because, until it is filled, a molecular understanding of the processes that determine disease progression will remain largely illusive. The long-term goal is to understand the pathways of tau misfolding and the processes of interneuronal spreading. The objective of this proposal is to use newly developed seeding methodology in conjunction with double electron-electron resonance (DEER) spectroscopy (a method that measures the distances between two paramagnetic reporter molecules) to determine the conformations of polypeptide chains in tau filaments. The central hypothesis is that conformationally distinct tau filaments are associated with different disease phenotypes. This hypothesis has been formulated based on preliminary data produced in the applicant's lab that reveal a robust seeding barrier between distinct isoforms of tau and structural polymorphism of filaments composed of identical isoforms. The rationale for this project is that once differences in filament structure are known, the effects of distinct conformers on cell transfer mechanisms can be tested, and the inhibition of these processes can be investigated. Supported by strong preliminary data the central hypothesis will be tested by pursuit of the following three specific aims. 1) Determine the seeding properties of tau filaments. A novel acrylodan-based fluorescence assay will be used to investigate the seeding properties of synthetically and brain-derived tau filaments and to characterize a newly discovered seeding barrier between tau isoforms. 2) Identify conformational differences between tau filaments. DEER spectroscopy and seeded filament growth will be used to determine conformational differences between tau filaments at the molecular level. 3) Determine filament properties of phosphorylated tau and tau disease mutants. The effects of phosphorylations and disease-related mutations on filament conformation will be investigated. Changes in seeding barriers between tau isoforms will serve to identify structural changes. Preliminary data have identified a mutation that causes a switch in seeding properties. Filaments of this mutant will be analyzed in detail using distance measurements between paramagnetic reporter groups. The research is innovative, because it uses a new, sensitive approach to resolve structural differences between filaments, namely conformational templating combined with double electron-electron resonance spectroscopy. The proposed research is significant, because the results will provide a first general model of filament diversity and its relationship to human tauopathies. Such knowledge has the potential to lead to new therapeutic strategies, including the production of novel monoclonal antibodies and small molecule inhibitors that selectively interfere with specific propagation routes, slowing or reversing the progression of tau-mediated diseases.

描述(由申请人提供)：在理解tau的错误折叠如何与在人类tauopathy中观察到的巨大表型多样性有关方面存在着根本的差距。这一差距的持续存在构成了一个重要的问题，因为在它被填补之前，对决定疾病进展的过程的分子理解在很大程度上仍将是虚幻的。长期目标是了解tau错误折叠的途径和神经元间扩散的过程。这项建议的目的是将新开发的种子方法与双电子-电子共振(DER)光谱(一种测量两个顺磁报告分子之间距离的方法)结合使用，以确定tau细丝中多肽链的构象。中心假设是构象不同的tau丝与不同的疾病表型有关。这一假设是基于申请人实验室产生的初步数据提出的，这些数据显示，不同的tau亚型和由相同亚型组成的细丝的结构多态之间存在强大的种子障碍。这个项目的基本原理是，一旦知道细丝结构的差异，就可以测试不同构象对细胞转移机制的影响，并可以研究这些过程的抑制。在强劲的初步数据的支持下，中心假说将通过追求以下三个具体目标来检验。1)确定Tau纤维的播种性能。一种新的基于acryloan的荧光分析将被用于研究合成的和脑来源的tau丝的种子特性，并表征新发现的tau亚型之间的种子屏障。2)确定tau丝之间的构象差异。鹿光谱和种子纤维生长将被用来在分子水平上确定tau纤维之间的构象差异。3)测定磷酸化tau和tau病突变体的细丝特性。将研究磷酸化和疾病相关突变对细丝构象的影响。Tau亚型之间播种障碍的变化将有助于识别结构变化。初步数据已经确定了一种突变，它会导致播种特性的改变。将使用顺磁报告基团之间的距离测量来详细分析该突变体的细丝。这项研究具有创新性，因为它使用了一种新的、灵敏的方法来解决细丝之间的结构差异，即构象模板结合双电子-电子共振光谱。这项拟议的研究意义重大，因为这些结果将提供第一个关于细丝多样性及其与人类自发性疾病的关系的通用模型。这些知识有可能导致新的治疗策略，包括生产新的单抗和小分子抑制剂，选择性地干扰特定的传播途径，减缓或逆转tau介导的疾病的进展。