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 阿尔茨海默氏病（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 微管相关的蛋白质TAU细胞内神经原纤维缠结（NFTS），细胞丧失和认知能力下降。 NFTS与临床症状的发作显着相关，但是当前调整疾病的疗法却没有 目标tau聚集机制。致病性错误折叠的tau在AD和 引起正常tau蛋白的“功能获得”错误折叠的方式。我们小组的证据 其他人已经确定了与AD中的病理TAU共同聚集的特定RNA结合蛋白（RBP） 大脑。这些RBP中的大多数是不溶性，将其定位在细胞质上，并显示剪接功能的损失 在广告中。重要的是，特定类别的RBP与AD病理增加和认知能力下降相关的RBP AD包含富含丝氨酸和精氨酸残基的低复杂性（LC）结构结构域 （SR-RICH）。目前尚不清楚富含SR的RBP是否改变了疾病中的Tau聚集。我建议这类 RBP通过其富含SR的LC域与AD中的TAU具有不同的相互作用。我的中心假设是 RBP与AD中的Tau共聚集将通过富含SR的LC结构域增强播种并增强Tau聚集。我会用 生化，细胞和体外测定的组合，以检查富含SR的LC结构域如何改变TAU 聚合。该项目的长期目标是通过调节tau来识别热干预点 AD的聚集并增强了我们对聚集过程的机械理解。目标1意志 确定RBP是否通过与Tau荧光的共定位通过富含SR的LC结构域介导Tau聚集 报告基细胞和硫非黄酮-T纤维化实验。在AIM 2中，我将通过一个 哺乳动物细胞中的新型Tau接近标记系统。该蛋白质组学屏幕提供了解决的能力 通过质谱技术和随后的生物信息学分析，其他共同聚集的RBP。 这两个平行目的将使我能够确定tau播种和聚集的机制 疾病。通过了解AD中RBP的SR丰富LC域如何促进Tau聚合，我将 有助于理解AD病理蛋白，并为治疗提供有影响力的见解 降低tau burnen的目标。通过成功完成提案AIM提供的培训将 重要的是我为神经退行性研究人员的独立研究人员做好了准备。