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: 10537598
• 负责人: Sarah Shapley
• 金额: $ 4.68万
• 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/10537598
• 关键词: Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Amyloid beta-Protein; Arginine; Binding; Biochemical; Bioinformatics; Biological Assay; Brain; 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; Lead; Length; MAPT gene; Mammalian Cell; Mass Spectrum Analysis; Mediating; Mediator of activation protein; 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; Seeds; Senile Plaques; Serine; Severities; Solubility; Structure; Symptoms; System; Tauopathies; Techniques; Testing; Therapeutic Intervention; Training; Transmission Electron Microscopy; United States; Western Blotting; base; 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.

项目总结 阿尔茨海默病(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， 微管相关蛋白tau细胞内神经原纤维缠结(NFTs)、细胞丢失和认知能力下降。 NFTs与临床症状的出现显著相关，但目前的疾病修正疗法并不是这样。 目标是tau聚集机制。致病错误折叠的tau在AD和AD的神经细胞中传播 导致正常tau蛋白以模板化方式错误折叠。来自我们小组的证据 另一些人发现了与AD病理性tau共同聚集的特定RNA结合蛋白(RBPs) 大脑。这些限制性商业惯例中的大多数是不能溶解的，错误地定位于细胞质，表现为剪接功能的丧失 在公元后。重要的是，一类特定的限制性商业惯例与AD病理增加和认知能力下降有关 AD含有无序的低复杂性(LC)结构域，富含丝氨酸和精氨酸残基 (富锶)。目前尚不清楚富含SR的限制性商业惯例是否会改变疾病中的tau聚集。我建议这个班级的 RBPs通过其富含SR的LC结构域与AD中的tau有明显的相互作用。我的中心假设是 在AD中，RBPs与tau共聚集将通过富含SR的LC结构域播种和增强tau聚集。我会用 生化、细胞和体外分析相结合的方法研究富含SR的LC结构域如何改变tau 聚合。这个项目的长期目标是通过调节tau来确定治疗干预点。 AD中的聚合，并增强我们对聚合过程的机械性理解。目标1将 确定限制性商业惯例是否通过与tau荧光共定位通过富含SR的LC结构域介导tau聚集 报告细胞和硫代黄素-T纤化实验。在目标2中，我将通过以下方式确定共同聚合的限制性商业惯例 哺乳动物细胞中新的tau邻近标记系统。这一蛋白质组筛选提供了解析 通过质谱学技术和随后的生物信息学分析进行额外的共聚合限制性商业惯例。 这两个平行的目标将使我能够确定tau播种和聚集的基础机制 疾病。通过了解AD中涉及的RBPs富含SR的LC结构域如何促进tau聚集，我将 有助于理解AD病理蛋白，并为治疗提供有效的见解 目标是减轻Tau负担。通过成功完成建议书目标提供的培训将 为我的职业生涯做好准备，成为一名神经退行性疾病的独立研究员。