Investigating the role of Serine Arginine (SR)-Rich RNA Binding Proteins in Tau Aggregation

研究富含丝氨酸精氨酸 (SR) 的 RNA 结合蛋白在 Tau 聚集中的作用

• 批准号: 10710487
• 负责人: Sarah Shapley
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10710487
• 关键词: Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Amyloid beta-Protein; Arginine; Binding; Biochemical; Bioinformatics; Biological Assay; Biotinylation; Brain; Cell Communication; Cell Culture Techniques; Cell Fractionation; Cell model; Cells; Chronic; Clinical; Cytoplasm; Dementia; Deposition; Detergents; Disease; Electron Transport; Embryo; Goals; Human; Image; Impaired cognition; Intervention; Kidney; Kinetics; Label; Length; MAPT gene; Mammalian Cell; Mass Spectrum Analysis; Mediating; Mediator; Microtubules; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Pathogenicity; Pathologic; Pathology; Pathway Analysis; Pathway interactions; Persons; Play; Process; Proteins; Proteome; Proteomics; RNA; RNA Recognition Motif; RNA Splicing; RNA-Binding Proteins; Reporter; Research; Research Personnel; Role; Senile Plaques; Serine; Severities; Solubility; Structure; Symptoms; System; Tauopathies; Techniques; Testing; Therapeutic Intervention; Training; Transmission Electron Microscopy; United States; Western Blotting; brain tissue; career; effective therapy; enhancing factor; experimental study; extracellular; gain of function; in vitro Assay; insight; loss of function; neurofibrillary tangle formation; new therapeutic target; novel; overexpression; paired helical filament; protein aggregation; recombinant RNA; self assembly; targeted treatment; tau Proteins; tau aggregation; therapeutic target

PROJECT SUMMARY Alzheimer’s disease (AD) is a chronic neurodegenerative disease characterized by extracellular β-amyloid (Aβ), microtubule-associated protein tau intracellular neurofibrillary tangles (NFTs), cellular loss, and cognitive decline. NFTs correlate significantly to the onset of clinical symptoms, yet current disease-modifying therapies do not target tau aggregation mechanisms. Pathogenic misfolded tau propagates across neuronal cells in AD and causes a ‘gain-of-function’ misfolding of normal tau proteins in a templated manner. Evidence from our group and others has identified specific RNA-binding proteins (RBPs) that co-aggregate with pathological tau in AD brains. A majority of these RBPs are insoluble, mislocalized to the cytoplasm, and display loss of splicing function in AD. Importantly, a specific class of RBPs that correlate with increased AD pathology and cognitive decline in AD contain disordered low complexity (LC) structural domains that are rich with serine and arginine residues (SR-rich). It is currently unknown if SR-rich RBPs alter tau aggregation in disease. I propose that this class of RBPs has distinct interactions with tau in AD through their SR-rich LC domains. My central hypothesis is that RBPs co-aggregating with tau in AD will seed and enhance tau aggregation via the SR-rich LC domain. I will use a combination of biochemical, cellular, and in vitro assays to examine how SR-rich LC domains alter tau aggregation. The long-term goal of this project is to identify points of therapeutic intervention by modulating tau aggregation in AD and enhancing our mechanistic understanding of the aggregation process. Aim 1 will determine if RBPs mediate tau aggregation via SR-rich LC domains through co-localization with tau fluorescent reporter cells and thioflavin-T fibrillization experiments. In Aim 2, I will identify co-aggregating RBPs through a novel tau proximity labeling system in mammalian cells. This proteomic screen provides the ability to resolve additional co-aggregating RBPs through mass spectrometry techniques and subsequent bioinformatic analyses. These two parallel aims will allow me to determine mechanisms that underlie tau seeding and aggregation in disease. By understanding how the SR-rich LC domain of RBPs implicated in AD promote tau aggregation, I will contribute to the understanding of AD pathological proteins and provide impactful insight toward therapeutic targets to reduce tau burden. The training provided through successful completion of the proposal aims will significantly prepare me for a career as an independent researcher in neurodegeneration.

项目摘要 阿尔茨海默病（Alzheimer's disease，AD）是一种以细胞外β-淀粉样蛋白（Aβ）为特征的慢性神经退行性疾病， 微管相关蛋白tau细胞内神经元缠结（NFT）、细胞损失和认知衰退。 NFT与临床症状的发作显著相关，但目前的疾病缓解疗法不 靶向tau聚集机制。致病性错误折叠的tau蛋白在AD中跨神经元细胞传播， 导致正常tau蛋白以模板方式的“功能获得性”错误折叠。我们小组的证据 等人已经鉴定出与AD中的病理性tau共聚集的特异性RNA结合蛋白（RBP 大脑这些RBP中的大多数是不溶性的，错误定位于细胞质，并显示剪接功能丧失 在AD中。重要的是，与AD病理学增加和认知功能下降相关的一类特定的RBP， AD含有富含丝氨酸和精氨酸残基的无序低复杂性（LC）结构域 （SR丰富）。目前尚不清楚富含SR的RBP是否会改变疾病中的tau聚集。我建议这类 RBP通过其富含SR的LC结构域与AD中的tau具有不同的相互作用。我的核心假设是 与AD中的tau共聚集的RBP将通过富含SR的LC结构域接种并增强tau聚集。我会用 结合生物化学、细胞和体外试验，研究富含SR的LC结构域如何改变tau蛋白 聚合来该项目的长期目标是通过调节tau蛋白来确定治疗干预点 聚集在AD和提高我们的机械理解的聚集过程。目标1将 确定RBP是否通过富含SR的LC结构域通过与tau荧光共定位介导tau聚集 报道细胞和硫磺素-T灭活实验。在目标2中，我将通过一个 哺乳动物细胞中的新型tau邻近标记系统。这种蛋白质组学筛选提供了解决 通过质谱技术和随后的生物信息学分析，对额外的共聚集限制性商业惯例进行分析。 这两个平行的目标将使我能够确定tau蛋白播种和聚集的基础机制， 疾病通过了解AD中涉及的RBP的富含SR的LC结构域如何促进tau聚集， 有助于了解AD病理蛋白，并为治疗提供有影响力的见解 目标是减少tau负担。通过成功完成提案目标提供的培训将 为我作为神经退行性疾病独立研究人员的职业生涯做好了重要准备。