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FUS Protein Homeostasis in ALS

ALS 中的 FUS 蛋白稳态

基本信息

批准号：
10550115
负责人：
Haining Zhu
金额：
--
依托单位：
SOUTHERN ARIZONA VA HEALTH CARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-10-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10550115
关键词：
ALS patients Address Affect Agonist Amyotrophic Lateral Sclerosis Animal Model Attenuated Bioinformatics Categories Cell Line Cell Nucleus Cell physiology Cells Cessation of life Contracts Cytoplasm Cytoplasmic Granules DNA Binding Defect Diagnosis Disease Drosophila genus Endoplasmic Reticulum Equilibrium Foundations Functional disorder Future Genes Genetic Genetic Models Health Healthcare Hydrogels Hyperactivity Impairment In Vitro Inclusion Bodies Intervention Knock-out Knowledge Liquid substance Mediating Membrane Proteins Messenger RNA Modeling Molecular Motor Neurons Mutation N-terminal Nerve Degeneration Neurodegenerative Disorders Neurons Nonsense-Mediated Decay Pathologic Pathology Pathway interactions Pharmacology Phase Phenotype Physiological Play Post-Translational Protein Processing Property Protein Biosynthesis Proteins Proteome Proteomics Publishing RNA RNA Binding RNA Decay RNA-Binding Proteins Rattus Reporting Ribonucleoproteins Role Services Specificity Testing Toxic effect Transgenic Mice Transgenic Organisms Translations Veterans amyotrophic lateral sclerosis therapy attenuation base care costs copper zinc superoxide dismutase cost design disability effective therapy familial amyotrophic lateral sclerosis fused in sarcoma high risk improved in vivo in vivo Model induced pluripotent stem cell inhibitor innovation insight mRNA Decay mRNA Surveillance military service military veteran mutant neuron loss neurotoxicity novel overexpression prion-like protein TDP-43 proteostasis sporadic amyotrophic lateral sclerosis tau Proteins therapeutic development transcriptome transcriptomics unpublished works

项目摘要

Amyotrophic lateral sclerosis (ALS) is a poorly understood neurodegenerative disease with no effective treatment. ALS is connected with military service and veterans are at a higher risk of developing this debilitating disease. Mutations in several genes have been discovered to cause a subset of familial ALS, including copper- zinc superoxide dismutase (SOD1), Fused in Sarcoma (FUS), and TAR DNA-binding protein 43 (TDP-43). Studying familial ALS using the well-defined genetic models will provide critical insights into the disease pathology as well as novel targets for future therapeutic development. This project is focused on the RNA binding protein FUS that has been implicated in both familial and sporadic ALS. ALS-related mutations in FUS cause a liquid-liquid phase separation (LLPS) of the FUS protein, forming liquid droplets in vitro. In neurons, mutant FUS accumulates in the cytoplasm, forming ribonucleoprotein granules and inclusions, eventually leading to neurotoxicity. We recently identified cellular proteins in FUS- positive inclusions and a bioinformatics analysis revealed two previously unknown pathways affected by mutant FUS: protein translation and mRNA surveillance. We also demonstrated that ALS FUS mutations indeed suppressed protein translation and hyper-activated the nonsense-mediated decay (NMD) of mRNAs. Our overarching hypothesis is that the dysregulation of protein translation and mRNA nonsense-mediated decay contributes to FUS toxicity and motor neuron dysfunction. Since little is known about how the mutant FUS- induced dysregulation of protein translation and NMD cause toxicity in neurons, discoveries in this project will fill this knowledge gap and advance the field significantly. Three specific aims are designed to test the hypothesis. Aim 1 is to determine what properties of mutant FUS drive the dysregulation of protein translation and NMD. We will use different domain truncations of FUS to manipulate LLPS and examine the relationship between LLPS and mutant FUS dysregulation. In addition, we will manipulate RNA binding of FUS and test whether RNA binding is a significant contributor to mutant FUS dysfunction. We will also use FUS knockout models to examine whether FUS plays a role in in protein translation and NMD under physiological conditions. Aim 2 is to determine whether mutant FUS impairs translation of specific proteins and NMD of specific mRNAs. We will employ specialized proteomic and transcriptomic approaches to identify changes of nascent protein biosynthesis and mRNA turnover rate as a consequence of mutant FUS. Differentially altered proteins and mRNAs will be validated in animal models as well as iPSC and induced motor neurons derived from familial ALS patients. Results from these -omics approaches will be integrated to determine whether mutant FUS impairs specific molecular and cellular function and pathways. Aim 3 is to examine whether attenuation of NMD hyperactivity can restore protein translation and mitigate FUS neurotoxicity. We will use genetic and pharmacological approaches to attenuate NMD hyperactivity and determine whether the intervention can restore the balance and mitigate FUS toxicity in vitro and in vivo models. The proposed studies are highly innovative, both conceptually and technically. The proposal is based on a number of novel observations regarding the impact of ALS mutant FUS on protein translation and mRNA decay. The results from this project are expected to produce new mechanistic insights into FUS ALS, laying a foundation for future therapeutic development of new ALS treatment(s).

肌萎缩侧索硬化症（ALS）是一种人们知之甚少的神经退行性疾病，目前尚无有效治疗方法治疗。 ALS 与服兵役有关，退伍军人患这种使人衰弱的疾病的风险更高疾病。已发现多个基因的突变可导致一部分家族性 ALS，其中包括铜- 锌超氧化物歧化酶 (SOD1)、肉瘤融合酶 (FUS) 和 TAR DNA 结合蛋白 43 (TDP-43)。使用明确的遗传模型研究家族性 ALS 将为对该疾病提供重要的见解病理学以及未来治疗发展的新靶点。该项目的重点是 RNA 结合蛋白 FUS，该蛋白与家族性和散发性 ALS。 FUS 中与 ALS 相关的突变会导致 FUS 蛋白发生液-液相分离 (LLPS)，在体外形成液滴。在神经元中，突变型 FUS 在细胞质中积累，形成核糖核蛋白颗粒和夹杂物，最终导致神经毒性。我们最近鉴定了 FUS 中的细胞蛋白阳性内含物和生物信息学分析揭示了受突变体影响的两条先前未知的途径 FUS：蛋白质翻译和 mRNA 监测。我们还证明 ALS FUS 突变确实抑制蛋白质翻译并过度激活 mRNA 的无义介导的衰变 (NMD)。我们的总体假设是蛋白质翻译失调和 mRNA 无义介导的衰变导致 FUS 毒性和运动神经元功能障碍。由于人们对突变体 FUS 的机制知之甚少，诱导的蛋白质翻译失调和 NMD 会导致神经元毒性，该项目的发现将填补这一知识差距并显着推进了该领域的发展。设计了三个具体目标来检验该假设。目标 1 是确定突变体的哪些特性 FUS 导致蛋白质翻译和 NMD 失调。我们将使用 FUS 的不同域截断来操纵 LLPS 并检查 LLPS 和突变型 FUS 失调之间的关系。此外，我们将操纵 FUS 的 RNA 结合并测试 RNA 结合是否是突变 FUS 的重要贡献者功能障碍。我们还将使用 FUS 敲除模型来检查 FUS 是否在蛋白质翻译中发挥作用和生理条件下的NMD。目标 2 是确定突变 FUS 是否会损害特定蛋白质和特定 mRNA 的 NMD。我们将采用专门的蛋白质组学和转录组学识别新生蛋白质生物合成和 mRNA 周转率变化的方法突变FUS。差异改变的蛋白质和 mRNA 将在动物模型以及 iPSC 和来自家族性 ALS 患者的诱导运动神经元。这些组学方法的结果将是整合以确定突变 FUS 是否损害特定的分子和细胞功能和途径。目的 3是检查NMD过度活跃的减弱是否可以恢复蛋白质翻译并减轻FUS 神经毒性。我们将使用遗传和药理学方法来减轻 NMD 过度活跃和确定干预措施是否可以在体外和体内模型中恢复平衡并减轻 FUS 毒性。拟议的研究在概念和技术上都具有高度创新性。该提案基于关于 ALS 突变体 FUS 对蛋白质翻译和 mRNA 影响的许多新观察衰变。该项目的结果预计将为 FUS ALS 提供新的机制见解，奠定为未来新的 ALS 治疗方法的开发奠定了基础。