Functional and pathological interactions of TDP-43

TDP-43 的功能和病理相互作用

• 批准号: 10133173
• 负责人: Nicolas Lux Fawzi
• 金额: $ 66.35万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10133173
• 关键词: Age; Aging; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amyotrophic Lateral Sclerosis; Binding; Binding Sites; Biochemical; Biological Assay; Biophysics; C-terminal; Cell Aggregation; Cell Nucleus; Cell model; Cell physiology; Cells; Cellular biology; Characteristics; Chemicals; Collaborations; Complex; Cyclophilin A; Cytoplasmic Granules; Data; Deposition; Disease; Disease model; Encephalopathies; Frontotemporal Dementia; Frontotemporal Lobar Degenerations; Future; Gene Mutation; Goals; Homeostasis; Human; Hydrogen Bonding; Hydrophobicity; Importins; Inclusion Bodies; Link; Liquid substance; Macromolecular Complexes; Maps; Mediating; Missense Mutation; Modification; Molecular; Molecular Biology; Molecular Probes; Motor Neuron Disease; Mutation; N-terminal; NMR Spectroscopy; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear Import; Nuclear Localization Signal; Pathogenicity; Pathologic; Pathology; Patients; Phase; Physiological; Poly Adenosine Diphosphate Ribose; Post-Translational Protein Processing; Process; Proteins; RNA Binding; RNA Processing; RNA Splicing; RNA-Binding Proteins; Research; Research Personnel; Resolution; Role; Structure; Techniques; Testing; Variant; age related; amyotrophic lateral sclerosis therapy; base; biophysical techniques; design; dimer; effective therapy; frontotemporal lobar dementia-amyotrophic lateral sclerosis; insight; limbic-predominant age-related TDP-43 encephalopathy; molecular modeling; novel; novel therapeutics; prevent; protein TDP-43; protein aggregation; simulation; structural biology; therapeutic development; therapeutic target; tool

Project Summary Cellular inclusions of proteins are primary hallmarks of a great majority of neurodegenerative diseases. In common forms of amyotrophic lateral sclerosis (ALS), Alzheimer’s disease related dementias (frontotemporal dementia; limbic-predominant age-related TDP-43 encephalopathy (LATE)) as well as some forms of Alzheimer’s disease, the essential human TAR DNA binding protein of 43 kDa (TDP-43) forms intraneuronal aggregates. Importantly, dozens of missense mutations in an aggregation-prone domain of TDP-43 have been found in familial and sporadic cases of ALS and frontotemporal dementia. These data provide strong support for the direct causative pathological role for TDP-43 in neurodegeneration in Alzheimer’s disease related dementias and motor neuron disease. Additionally, recent research modulating the TDP-43 interactome demonstrates that TDP-43 is an important potential therapeutic targets in these diseases. However, therapeutic development is hampered in large part by an absence of mechanistic understanding of normal TDP-43 function, the molecular effect of the mutations causing amyotrophic lateral sclerosis and frontotemporal dementia, and the TDP-43 disruption in neurodegenerative disease. These gaps are due in large part to a lack of atomic structural data regarding TDP-43, its complexes, and its conversion to aggregates, which in turn is due to the difficulty in observing TDP-43 complexes via traditional structural biology techniques. This project will make use of integrated experimental and computational structural biology techniques combined with molecular and cell biology approaches to 1) determine the atomistic details of the assembly of a helical sub-region of TDP-43, its contribution to splicing function, and its structural conversion in disease aggregates, 2) identify how known and novel post-translational modifications and disease-associated mutations alter TDP-43 self- and hetero-protein contacts that mediate and regulate TDP-43 liquid-liquid phase separation and disease-associated aggregation, and 3) map the structural basis of the interactions of TDP-43 with poly(ADP-ribose) and importin machinery that serve as promising therapeutic targets. The challenging, dynamic, structural targets necessitate the approach highlighting a tight connection between molecular simulation and experimental biophysical techniques (primarily NMR spectroscopy). These approaches will generate detailed molecular models of the interactions that will be tested for functional relevance using in cell aggregation, in cell splicing, and in cell protein/RNA binding structure (iCLIP). The results of these studies on TDP-43 complexes, phase-separation, function, and aggregation will provide direct structural and mechanistic input to the design of strategies to prevent toxic disruption of TDP-43 in Alzheimer’s disease related dementias and motor neuron disease. These insights represent potential for future treatments for ALS, frontotemporal dementia, and other TDP-43-associated diseases that currently have no cure or effective treatments.

项目摘要 蛋白质的细胞内含物是绝大多数神经退行性疾病的主要标志。在 常见的肌萎缩侧索硬化症（ALS）、阿尔茨海默病相关痴呆（额颞 痴呆;边缘优势年龄相关性TDP-43脑病（LATE））以及某些形式的 阿尔茨海默病，43 kDa的人类TAR DNA结合蛋白（TDP-43）形成神经元内 集料.重要的是，TDP-43的聚集倾向结构域中的数十个错义突变已经被发现。 在ALS和额颞叶痴呆的家族性和散发性病例中发现。这些数据提供了有力的支持 TDP-43在阿尔茨海默病相关神经变性中的直接致病病理作用 痴呆和运动神经元疾病。此外，最近调节TDP-43相互作用组的研究 表明TDP-43是这些疾病中重要的潜在治疗靶点。然而，在这方面， 治疗的发展在很大程度上是由于缺乏对正常的 TDP-43的功能，导致肌萎缩侧索硬化症的突变的分子效应， 额颞叶痴呆和神经退行性疾病中的TDP-43破坏。这些差距是由于在 这在很大程度上是由于缺乏关于TDP-43、其复合物及其转化为 聚集体，这又是由于难以通过传统的结构分析观察TDP-43复合物。 生物技术这个项目将利用综合实验和计算结构生物学 技术结合分子和细胞生物学方法，以1）确定细胞的原子细节， TDP-43的螺旋亚区的组装，其对剪接功能的贡献，以及其在 疾病聚集体，2）确定已知的和新的翻译后修饰和疾病相关的 突变改变TDP-43自身和异源蛋白接触，介导和调节TDP-43液-液相 分离和疾病相关聚集，以及3）绘制TDP-43相互作用的结构基础 与聚（ADP-核糖）和输入机制，作为有前途的治疗目标。挑战， 动态的，结构的目标需要的方法强调分子之间的紧密联系， 模拟和实验生物物理技术（主要是NMR光谱）。这些办法将 生成相互作用的详细分子模型，这些模型将用于测试细胞内的功能相关性。 聚集、细胞剪接和细胞蛋白/RNA结合结构（iCLIP）。这些研究的结果， TDP-43复合物，相分离，功能和聚集将提供直接的结构和机制 为设计预防阿尔茨海默病相关痴呆中TDP-43毒性破坏的策略提供了投入 和运动神经元疾病。这些见解代表了未来ALS治疗的潜力，额颞叶 痴呆症和其他TDP-43相关疾病，目前没有治愈或有效的治疗方法。