Functional and pathological interactions of TDP-43

TDP-43 的功能和病理相互作用

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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.

项目概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。常见形式的肌萎缩侧索硬化症 (ALS)、阿尔茨海默病相关痴呆（额颞叶痴呆；边缘系统主导的年龄相关 TDP-43 脑病（LATE））以及某些形式的阿尔茨海默氏病，43 kDa 的必需人类 TAR DNA 结合蛋白 (TDP-43) 形成神经元内重要的是，TDP-43 的易聚集结构域中存在数十个错义突变。在 ALS 和额颞叶痴呆的家族性和散发性病例中发现的这些数据提供了强有力的支持。 TDP-43 在阿尔茨海默病相关神经变性中的直接致病作用此外，最近的研究还调节 TDP-43 相互作用组。表明 TDP-43 是这些疾病的重要潜在治疗靶点。由于缺乏对正常机制的理解，治疗的发展在很大程度上受到阻碍。 TDP-43 功能、导致肌萎缩侧索硬化症的突变的分子效应和额颞叶痴呆和神经退行性疾病中的 TDP-43 破坏。很大程度上是由于缺乏有关 TDP-43、其配合物及其转化的原子结构数据聚集体，这又是由于通过传统结构观察 TDP-43 复合物的困难该项目将利用综合实验和计算结构生物学。技术与分子和细胞生物学方法相结合，1) 确定 TDP-43螺旋子区域的组装、其对剪接功能的贡献及其结构转换疾病聚合体，2) 确定已知和新颖的翻译后修饰与疾病相关的程度突变改变 TDP-43 自身蛋白和异源蛋白接触，从而介导和调节 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聚（ADP-核糖）和导入机制是有前途的治疗靶标。动态的结构目标需要强调分子之间紧密联系的方法这些方法将采用模拟和实验生物物理技术（主要是核磁共振波谱）。生成详细的相互作用分子模型，将在细胞中测试其功能相关性这些研究的结果涉及细胞剪接和细胞蛋白质/RNA 结合结构 (iCLIP)。 TDP-43复合物、相分离、功能和聚集将提供直接的结构和机械性能为预防阿尔茨海默病相关痴呆中 TDP-43 的毒性破坏策略的设计提供参考这些见解代表了 ALS、额颞叶疾病的未来治疗的潜力。痴呆症和其他目前尚无治愈或有效治疗方法的 TDP-43 相关疾病。