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.

肌萎缩性侧索硬化症（ALS）是一种进行性神经退行性疾病，其中首选的损失运动神经元（MNS）导致瘫痪和死亡。尽管ALS在很大程度上是一种零星疾病，但研究已经专注于可遗传的疾病形式，因为临床和病理证据表明常见致病机制。基因FUS中的突变引起了一些最激进的早期发作形式 ALS。 FUS病理学 - 很少存在突变 - 也与相关的神经退行性疾病有关额颞痴呆（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体 FU会导致运动神经元变性，而不是通过功能丧失，而有毒的功能获得不涉及过量的毒气活性。 FUS是许多RNA结合蛋白之一 - 包括TDP-43和HNRNP A1 - 偶尔与ALS和FTD有关。最近的工作导致了疾病模型 FUS和相关蛋白质本质上无序的“风雨状”结构域驱动相变，导致导致不可逆的神经毒性骨料的形成。 FUS中与ALS相关的突变会增加自然蛋白质形成这些有毒组件的趋势，这些组件会捕获和隔离其他核糖核蛋白颗粒成分。在这个项目中，我们将探索一系列新型的敲入突变体中FUS毒性的机制重现FUS-ALS表型的关键方面的小鼠。此外，使用运动神经元的体外研究从这些小鼠模型得出的星形胶质细胞将用于研究细胞自主和非 - 疾病的自主机制。在AIM 1中，我们将使用条件敲击鼠标模型表达 MNS或星形胶质细胞中的突变FU，或在神经系统中更广泛地探索暂时的影响并在MN的存活和功能上在空间调节突变型FUS表达；我们还将探索亲戚人体FUS的毒性在完全人性化的FUS-ALS小鼠模型中。在这个高度疾病的模型中 ALS/FTD，我们还将测试FUS脱聚类酶的治疗潜力，Kap2作为减速或停止的一种手段 MN变性的开始和进展。在AIM 2中，我们将结合精致的电生理学和探索在整个疾病进展中突变FUS的功能后果的行为方法 FUS敲入鼠标。最后，在AIM 3中，我们将使用单细胞RNA测序和拓扑结合数据分析到从我们的FUS敲入突变小鼠中得出的体外分化MN的混合分布。体内和体外实验系统的这种复杂整合，并结合我们的整合计算和分析方法将使我们能够阐明易受伤害的疾病途径 MN的亚群，并确定用于治疗FUS-ALS及其相关的潜在治疗靶标运动神经元疾病的形式。