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

• 批准号: 8338799
• 负责人: Martin Margittai
• 金额: $ 30.3万
• 依托单位: UNIVERSITY OF DENVER (COLORADO SEMINARY)
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8338799
• 关键词: Adult; Alzheimer' 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; 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

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tau蛋白病中观察到的巨大表型多样性相关方面存在根本性差距。这个缺口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将在很大程度上仍然是虚幻的。长期目标是了解tau蛋白错误折叠的途径和神经元间扩散的过程。该提案的目的是使用新开发的接种方法结合双电子-电子共振（DEER）光谱（一种测量两个顺磁性报告分子之间距离的方法）来确定tau蛋白丝中多肽链的构象。中心假设是构象不同的tau丝与不同的疾病表型相关。该假设是基于申请人实验室中产生的初步数据制定的，这些数据揭示了tau的不同同种型和由相同同种型组成的细丝的结构多态性之间的稳健播种屏障。该项目的基本原理是，一旦已知细丝结构的差异，就可以测试不同构象对细胞转移机制的影响，并研究这些过程的抑制。在强有力的初步数据的支持下，将通过追求以下三个具体目标来检验中心假设。1)确定tau纤维的接种特性。一种新的基于丙烯酰胺的荧光测定将用于研究合成和脑衍生的tau细丝的播种特性，并表征tau亚型之间新发现的播种屏障。2)识别tau蛋白丝之间的构象差异。DEER光谱和接种丝生长将用于在分子水平上确定tau丝之间的构象差异。3)确定磷酸化tau蛋白和tau蛋白疾病突变体的纤维特性。将研究磷酸化和疾病相关突变对细丝构象的影响。tau亚型之间接种屏障的变化将用于鉴定结构变化。初步数据已经确定了一种导致播种特性转换的突变。将使用顺磁性报告基团之间的距离测量来详细分析该突变体的细丝。这项研究是创新的，因为它使用了一种新的，敏感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。这项研究具有重要意义，因为研究结果将提供第一个细丝多样性及其与人类tau蛋白病关系的一般模型。这些知识有可能导致新的治疗策略，包括生产新的单克隆抗体和小分子抑制剂，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。