Patient-Based Structural Biology of Tauopathies and TDP-43 Proteinopathies using Cryo-Electron Microscopy and Mass Spectrometry

使用冷冻电子显微镜和质谱法对 Tau 蛋白病和 TDP-43 蛋白病进行基于患者的结构生物学研究

• 批准号: 10011921
• 负责人: Anthony William Paul Fitzpatrick
• 金额: $ 162.92万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 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 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011921
• 关键词: Affinity; Alzheimer' s Disease; Alzheimer' s disease brain; Amyotrophic Lateral Sclerosis; Animals; Antibodies; Autopsy; Behavioral; Behavioral Symptoms; Biochemical; Biological Markers; C-terminal; Cell model; Cells; Clinical; Complex; Cryoelectron Microscopy; Cytoplasmic Inclusion; Data; Dementia; Deposition; Development; Disease; Event; Exhibits; Filament; Frontotemporal Lobar Degenerations; Generations; Goals; Heterogeneity; Immunotherapy; In Vitro; Knowledge; Ligands; Link; Lobar; Mass Spectrum Analysis; Mediating; Methodology; Modeling; Modification; Molecular Biology; Morphology; Mutation; Nerve Degeneration; Neurites; Neurobehavioral Manifestations; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Phenotype; Phosphorylation; Positron-Emission Tomography; Post-Translational Protein Processing; Progressive Supranuclear Palsy; Property; Protein Isoforms; Reagent; Recombinants; Resource Development; Resources; Role; Structure; Tauopathies; Tissues; Toxic effect; Ubiquitination; base; brain tissue; chronic traumatic encephalopathy; cognitive function; in vivo; insight; motor symptom; mutation carrier; novel; novel diagnostics; novel therapeutics; protein TDP-43; small molecule; structural biology; tau Proteins; tau aggregation; tau phosphorylation; tool

The development of an atomic model of tau filaments in Alzheimer’s disease (AD) has opened several new opportunities, including the ability to identify high-affinity ligands or antibodies to detect AD tau filaments (i.e., PET ligands, small molecule compounds, immunotherapy, biomarkers) and providing a clear directive about essential features that need to be recapitulated from a modeling perspective. Given that tauopathies are complex, pathogenic entities with significant heterogeneity in clinical presentation, disease course, and underlying pathology, it is essential that the tools under development are capable of detecting/recognizing the actual pathologies observed. As such, the overall goals of the current project are to characterize the heterogeneity of tau filaments across tauopathies, and to utilize structural features that define tau and TDP-43 filament morphology in sporadic and familial forms of disease to provide novel insight into pathogenesis and toxicity. In particular, we will assess and compare tau filament heterogeneity between AD and chronic traumatic encephalopathy (CTE) and develop novel resources to both detect and model tau in these distinct tauopathies. Similar methodology utilizing cryo-electron microscopy (cryo-EM) will also be employed to resolve filament structure in frontotemporal lobar degeneration (FTLD), which is the second most common cause of dementia after AD. As FTLD is usually characterized pathologically by the aberrant deposition of either tau (FTLD-tau) or TDP-43 (FTLD-TDP), we will develop atomic models of both tau and TDP-43 filaments in sporadic and familial forms of FTLD. Overlaying structural insights with our recent discovery of novel post-translational modifications (PTMs) of tau and TDP-43, we will assess the contribution of specific modifications to aggregation propensity, utilizing a combination of cryo-EM, mass spectrometry, molecular biology and biochemical approaches. These results will guide the continued development of resources to best detect and model particular aspects of pathology.

阿尔茨海默病（AD）中tau蛋白丝原子模型的发展开辟了几个新的机会，包括鉴定高亲和力配体或抗体来检测AD tau蛋白丝（即PET配体、小分子化合物、免疫疗法、生物标志物）的能力，并提供了从建模角度需要概括的基本特征的明确指示。鉴于牛头病是复杂的致病实体，在临床表现、病程和潜在病理方面具有显著的异质性，因此开发的工具必须能够检测/识别观察到的实际病理。因此，当前项目的总体目标是表征tau丝在tau病中的异质性，并利用散发性和家族性疾病中定义tau和TDP-43丝形态的结构特征，为发病机制和毒性提供新的见解。特别是，我们将评估和比较AD和慢性创伤性脑病（CTE）之间的tau丝异质性，并开发新的资源来检测和模拟这些不同的tau病。利用冷冻电镜（cryo-EM）的类似方法也将被用于解决额颞叶变性（FTLD）的纤维结构，这是仅次于阿尔茨海默氏症的第二大痴呆原因。由于FTLD的病理特征通常是tau （FTLD-tau）或TDP-43 （FTLD- tdp）的异常沉积，我们将在散发性和家族性FTLD中建立tau和TDP-43细丝的原子模型。结合我们最近发现的tau和TDP-43的新型翻译后修饰（PTMs）的结构见解，我们将利用冷冻电镜、质谱、分子生物学和生化方法的结合，评估特定修饰对聚集倾向的贡献。这些结果将指导资源的持续发展，以最好地检测和模拟病理的特定方面。