Defining The Impaired Proteostasis Network in ALS Patient Motor Neurons

定义 ALS 患者运动神经元受损的蛋白质稳态网络

基本信息

批准号：
9756483
负责人：
Evangelos Kiskinis
金额：
$ 19.75万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9756483
关键词：
Affect Amyotrophic Lateral Sclerosis Axonal Transport C9ORF72 Carrier Proteins Cell physiology Cells Cessation of life Characteristics Code Coupled DNA Sequence Alteration Data Defect Degradation Pathway Disease Etiology Event Exhibits Functional disorder Gene Mutation Genes Genetic Genetic study Homeostasis Human Impairment Intervention Light Link Literature Mass Spectrum Analysis Mediating Microscopic Mitochondria Molecular Motor Motor Neurons Movement Muscle Mutate Mutation Nature Nerve Degeneration Network-based Neurodegenerative Disorders Neurons Pathway interactions Patients Pattern Protein Analysis Protein Biosynthesis Proteins Proteome Proteomics Publishing Quality Control RNA Processing Regulation Reporting Research Resolution Resources Testing Therapeutic Intervention Time Translations Transport Vesicles Vesicle Transport Pathway Work base causal variant effective therapy experimental study gene correction genetic variant improved induced pluripotent stem cell innovation motor control motor neuron degeneration mutant nervous system disorder neurotoxic protein aggregate protein degradation protein folding protein transport proteostasis superoxide dismutase 1 therapeutic target

项目摘要

Amyotrophic lateral sclerosis (ALS) is a progressive and untreatable neurodegenerative disease that is characterized by the selective death of upper and lower motor neurons (MNs). The overwhelming majority of the disease is sporadic in nature. However a relatively small (<12%) but highly informative fraction of patients suffer from familial forms of disease, which have enabled the identification of causative genetic variants that underlie their condition. Such genetic studies have demonstrated that ALS can be caused by mutations in genes that encode proteins involved in diverse set of cellular functions ranging from RNA processing, vesicle transport, cytoskeletal regulation, mitochondrial function, and protein quality control pathways. Nevertheless, ALS patients are uniformly characterized by a common pattern of progressive motor neurodegeneration. This raises the possibility that different disease initiating events could coalesce in one or more common molecular pathways. How the mutation of genes with dissimilar functions converge on MN degeneration has been and continues to be an outstanding question. Although all ALS patients exhibit neuropathological protein aggregates, the overall contribution of protein homeostasis in causing ALS has remained unclear. If we could identify a convergent mechanism, it may provide an opportunity to develop a broadly applicable therapeutic intervention strategy. In our preliminary studies, we conducted global analysis of protein degradation dynamics in mutant SOD1 and isogenic controls MNs derived from iPSC lines. Interestingly, we identified a number of proteins that are degraded at a slower rate in SOD1 MNs. Unexpectedly, this small panel of candidates included proteins whose genetic mutations cause ALS. In the proposed research we will use patient-derived neurons coupled with mass spectrometry analysis to determine the protein substrates, as well as the nature of the perturbation that arise as a result of mutations in the two most prevalent ALS genes: SOD1 and C9orf72. First, we will determine which proteins have reduced protein degradation dynamics. Second, we will determine which proteins have altered synthesis rates. Third, we will determine the overall degree of proteome-wide remodeling. Each of these approaches has strategic advantages over traditional work-flows and will allow us to determine not only which proteins have altered levels in ALS MNs but also the mechanism responsible for their perturbation. Taken together, our proposed aims will shed light into the cellular mechanisms compromised by changes in the proteostasis network in patient neurons and will likely uncover broadly relevant therapeutic targets for ALS.

肌萎缩性侧索硬化症（ALS）是一种进行性且不可治疗的神经退行性疾病以上和下运动神经元（MN）的选择性死亡为特征。绝大多数这种疾病本质上是零星的。但是，患者的相对较小（<12％）但信息丰富患有家族形式的疾病，这使得能够鉴定出病因的遗传变异是他们的状况。这种遗传研究表明，ALS可能是由突变引起的编码参与各种细胞功能的蛋白质的基因，范围从RNA加工，囊泡运输，细胞骨架调节，线粒体功能和蛋白质质量控制途径。尽管如此， ALS患者的统一特征是进行性运动神经变性的常见模式。这提高了不同疾病启动事件可以在一个或多个共同的分子中合并的可能性途径。具有不同功能的基因突变在MN变性上的收敛性和仍然是一个重大的问题。尽管所有ALS患者均表现出神经病理蛋白聚集体，蛋白质稳态在引起ALS的总体贡献尚不清楚。如果可以的话确定收敛机制，它可能会提供开发广泛适用治疗的机会干预策略。在我们的初步研究中，我们对蛋白质降解动力学进行了全球分析在突变体SOD1和源自IPSC系的ISEGENIC CONTRORS MN中。有趣的是，我们确定了许多在SOD1 MN中以较慢的速率降解的蛋白质。出乎意料的是，这个小组的候选人面板包括其基因突变引起ALS的蛋白质。在拟议的研究中，我们将使用患者来源神经元与质谱分析结合以确定蛋白质底物以及由于两个最普遍的ALS基因突变而产生的扰动：SOD1和C9ORF72。首先，我们将确定哪些蛋白质减少了蛋白质降解动力学。第二，我们将确定哪些蛋白质改变了合成速率。第三，我们将确定全蛋白质组的整体程度重塑。这些方法中的每一种都比传统的工作流程具有战略优势，将使我们能够不仅确定哪些蛋白质在ALS MN中有所改变，还确定负责其水平的水平扰动。综上所述，我们提出的目标将使光明患者神经元中蛋白抑制网络的变化，可能会发现广泛相关的治疗性 ALS的目标。