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Mechanisms of FUS Toxicity in Animal and Cellular Models of ALS/FTD.

FUS 在 ALS/FTD 动物和细胞模型中的毒性机制。

基本信息

批准号：
10337336
负责人：
Neil Alan Shneider
金额：
$ 64.8万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-15 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10337336
关键词：
Action Potentials Adolescent Alleles Amyotrophic Lateral Sclerosis Animal Disease Models Animal Model Animals Astrocytes Behavior monitoring Behavioral Biological Models Cell Differentiation process Cell model Cells Cessation of life Clinical Cognitive Computing Methodologies Cytoplasmic Granules Data Analyses Defect Development Disease Disease Pathway Disease Progression Disease model Dose Electrophysiology (science)Evaluation Frontotemporal Dementia Functional disorder Gene Activation Genes Genetic Genetic Transcription Heritability Heterogeneity Hindlimb Human Immunohistochemistry In Vitro Karyopherins Knock-in Knock-in Mouse Lead Mediating Methodology Methods Modeling Molecular Molecular Chaperones Molecular Computations Morphology Motor Motor Neuron Disease Motor Neurons Mus Muscle Mutant Strains Mice Mutation Natural Increases Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neuroglia Neuronal Dysfunction Neurons Nuclear Nuclear Import Onset of illness Paralysed Pathogenicity Pathologic Pathology Patients Pattern Phase Phase Transition Phenotype Physiological Population Preparation Prion Diseases Proteins RNA-Binding Proteins Research Respiratory Diaphragm Ribonucleoproteins Series Spinal Cord System Testing Time Toxic effect Viral Viral Vector Work behavior measurement behavior test behavioral study cell type cognitive testing early onset embryonic stem cell executive function experimental study extracellular frontotemporal degeneration frontotemporal lobar dementia-amyotrophic lateral sclerosis gain of function hnRNP A1 humanized mouse in vivo insight loss of function motor behavior motor neuron degeneration mouse genetics mouse model mutant mutant mouse model neuromuscular activity neuromuscular function neuromuscular transmission neuron loss neurotoxic new therapeutic target novel novel therapeutics overexpression prevent prion-like protein TDP-43 receptor selective expression single cell sequencing single-cell RNA sequencing stress granule targeted treatment therapeutic development therapeutic evaluation therapeutic target

项目摘要

Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder in which preferential loss of motor neurons (MNs) results in paralysis and death. Although ALS is largely a sporadic disease, research has focused on heritable forms of the disorder because clinical and pathological evidence suggests common pathogenic mechanisms. Mutations in the gene FUS cause some of the most aggressive early-onset forms of ALS. FUS pathology – and rarely, mutations - are also associated with the related neurodegenerative disorder, frontotemporal dementia (FTD). In a recent study, our lab demonstrated in a mouse model of disease that mutant FUS causes motor neuron degeneration not by a loss-of-function, by a toxic gain-of-function that does not involve an excess of FUS activity. FUS is one of a number of RNA binding proteins – including TDP-43 and hnRNP A1 – that have been causally related to ALS and FTD. Recent work has led to a disease model in which the intrinsically disordered “prion-like” domain of FUS and related proteins drives a phase transition that results in the formation of an irreversible, neurotoxic aggregate. ALS-related mutations in FUS increase the natural tendency of the protein to form these toxic assemblies, which trap and sequester other ribonucleoprotein granule components. In this project we will explore the mechanisms of FUS toxicity in a novel series of knock-in mutant mice that reproduce key aspect of the FUS-ALS phenotype. In addition, in vitro studies using motor neurons and astrocytes derived from these mouse models will be used to investigate cell autonomous and non- autonomous mechanisms of disease. In Aim 1, we will use a conditional knock-in mouse model to express mutant FUS in MNs or astrocytes, or more broadly in the nervous system to explore the effects of temporally and spatially regulated mutant FUS expression on MN survival and function; and we will also explore the relative toxicity of human FUS in a fully humanized mouse model of FUS-ALS. In this highly disease-relevant model of ALS/FTD, we will also test the therapeutic potential of the FUS disaggregase, Kap2 as a means to slow or stop the onset and progression of MN degeneration. In Aim 2, we will combine sophisticated electrophysiological and behavioral methods to explore the functional consequence of mutant FUS throughout disease progression in the FUS knock-in mouse. Finally in Aim 3, we will apply a combination of single-cell RNA sequencing and topological data analysis to a mixed distribution of in vitro differentiated MNs derived from our FUS knock-in mutant mice. This sophisticated integration of in vivo and in vitro experimental systems, combined with our integrative computational and analytical approach will allow us to elucidate pathways of disease in vulnerable subpopulations of MNs and to identify potential therapeutic targets for the treatment of FUS-ALS and related forms of motor neuron disease.

肌萎缩侧索硬化症(ALS)是一种进行性神经退行性疾病，在这种疾病中运动神经元(MN)会导致瘫痪和死亡。尽管肌萎缩侧索硬化症在很大程度上是一种散发性疾病，但研究已经重点关注这种疾病的可遗传形式，因为临床和病理证据表明致病机制。FUS基因的突变导致了一些最具侵袭性的早发性肌萎缩侧索硬化。FUS病理--很少是突变--也与相关的神经退行性疾病有关，额颞性痴呆(FTD)。在最近的一项研究中，我们的实验室在疾病的小鼠模型中证明了突变 FUS导致运动神经元退化不是由于功能丧失，而是由于不涉及 FUS活动过多。FUS是多种RNA结合蛋白之一，包括TDP-43和hnRNP A1-与ALS和FTD有因果关系。最近的工作导致了一种疾病模型，在这种模型中， FUS和相关蛋白的固有无序的“类普里子”结构域驱动相变，导致形成不可逆的、神经毒性的聚集体。肌萎缩侧索硬化症相关基因突变增加了自然蛋白质形成这些有毒集合体的倾向，这些集合体捕获和隔离其他核糖核蛋白颗粒组件。在这个项目中，我们将探索一系列新的敲入突变体的FUS毒性机制。复制FUS-ALS表型关键方面的小鼠。此外，使用运动神经元进行的体外研究从这些小鼠模型中获得的星形胶质细胞将被用于研究细胞自主和非疾病的自主机制。在目标1中，我们将使用条件敲入小鼠模型来表达突变型FUS在MNS或星形胶质细胞中，或更广泛地在神经系统中探索暂时性的影响和空间调控突变体FUS的表达对MN存活和功能的影响；我们还将探讨其相对人FUS在完全人源化的FUS-ALS小鼠模型中的毒性。在这个与疾病高度相关的模型中，我们还将测试FUS解聚酶KAP2作为减缓或停止的手段的治疗潜力微核退行性变的发生和发展。在目标2中，我们将结合复杂的电生理学和用行为学方法探讨突变的FUS在疾病进展过程中的功能后果 Fus敲入鼠标。最后，在目标3中，我们将应用单细胞RNA测序和拓扑学的组合来自我们的FUS敲入突变小鼠的体外分化的MN混合分布的数据分析。这种体内和体外实验系统的复杂集成，与我们的集成计算和分析方法将使我们能够阐明脆弱人群的疾病途径 MN的亚群和寻找治疗FUS-ALS及相关疾病的潜在靶点运动神经元病的形式。