Investigating the impact of p97 mutation in Amyotrophic Lateral Sclerosis

研究 p97 突变对肌萎缩侧索硬化症的影响

基本信息

批准号：
10558598
负责人：
Malavika Raman
金额：
$ 20.63万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10558598
关键词：
ATP Hydrolysis ATP phosphohydrolase Adaptor Signaling Protein Affinity Affinity Chromatography Aging Amyotrophic Lateral Sclerosis Autophagocytosis Binding Brain Stem Cell Aging Cell Cycle Cell model Cell physiology Cells Cessation of life Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex Dedications Denervation Development Diagnosis Disease Disease Progression Ensure Enzymes Future Gene Mutation Generations Genes Glycylglycine Goals Healthcare Heterozygote Homeostasis Impairment Inclusion Body Myopathy with Early-Onset Paget Disease Individual Intervention Lead Left Link Missense Mutation Mitochondria Modeling Molecular Molecular Chaperones Motor Neuron Disease Motor Neurons Muscle Muscle Fibers Muscle function Muscular Atrophy Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Organelles Pathway interactions Patients Phenotype Physiology Prevalence Protein Biosynthesis Proteins Proteome Proteomics Quality Control Reporting Research Role Skeletal Muscle Specificity Spinal Cord Symptoms System Testing Transformed Cell Line Triage Ubiquitin Ubiquitination United States Variant age related neurodegeneration aging population bone cell age cell type cohort differentiation protocol disease phenotype familial amyotrophic lateral sclerosis induced pluripotent stem cell insight loss of function multicatalytic endopeptidase complex mutant nervous system disorder new therapeutic target non-Native overexpression prevent protein aggregation proteostasis small molecule stem cell model therapy development unfoldase

项目摘要

Protein homeostasis decline is a significant contributor to the development of Amyotrophic Lateral Sclerosis (ALS) and other neurodegenerative disorders. With an ever-expanding aging population and no reliable therapies, these devastating diseases will place a significant burden on health care in the coming decades. The p97 AAA-ATPase is a ubiquitin-selective unfoldase that has critical roles in protein quality control. Dedicated adaptor proteins target p97 to ubiquitylated substrates and thereby impart specificity to this abundant enzyme. Mutations in p97 cause familial ALS and are reported to impede p97 association with certain adaptors and impair ER and mitochondria associated quality control suggesting inadequate targeting to these organelles. However, the impact of p97 mutation on its interactome and ubiquitylated substrates has not been studied systematically in cell types impacted in disease, namely motor neurons and skeletal muscle. Here, we will establish an induced pluripotent stem cell (iPSC)-based model of motor neuron disease caused by distinct p97 mutations. We will use this system to: (1) Investigate how mutant p97 remodels the motor neuron and skeletal muscle proteome and physiology. Ubiquitylated substrates of p97 will be identified using quantitative proteomics in each cell type. (2) Examine the impact of p97 mutation on known protein quality control pathways in motor neurons skeletal muscle cells. Our studies will provide the first model of ALS caused by p97 mutation and will provide mechanistic insight into how p97 mutation causes motor neuron and/or skeletal muscle demise. We anticipate our cell-based model may be used in the future for small molecule screens to identify lead molecules for therapy development. Given the prevalence of p97 mutation, we anticipate these studies will have broad relevance to other in neurodegenerative diseases.

蛋白质稳态下降是肌萎缩性侧索硬化症发展的重要原因（ALS）和其他神经退行性疾病。人口不断扩大，没有可靠疗法，这些毁灭性的疾病将在未来几十年中给医疗保健带来重大负担。 p97 AAA-ATPase是一种泛素选择性展进期，在蛋白质质量控制中具有关键作用。专用的适配器蛋白靶标P97对泛素化的底物，从而赋予这种丰富的特异性酶。 p97的突变引起家族性ALS，并据报道会阻碍P97与某些适配器的关联损害ER和线粒体相关的质量控制表明对这些细胞器的靶向不足。但是，尚未研究p97突变对其相互作用组和泛素化底物的影响系统地在疾病影响的细胞类型中，即运动神经元和骨骼肌。在这里，我们将建立一个诱导的多能干细胞（IPSC）的运动神经元疾病模型通过不同的P97突变。我们将使用该系统来：（1）研究突变体P97如何重塑运动神经元以及骨骼肌蛋白质组和生理学。将使用定量确定p97的泛素化底物每种细胞类型中的蛋白质组学。（2）检查p97突变对已知蛋白质质量控制途径的影响在运动神经元中骨骼肌细胞。我们的研究将提供由P97突变引起的第一个ALS模型，并将提供机械洞察力 p97突变如何导致运动神经元和/或骨骼肌灭亡。我们预计我们的基于细胞的模型将来可以用于小分子筛选，以鉴定铅分子以进行治疗。给出 p97突变的流行率，我们预计这些研究将与其他研究具有广泛的相关性神经退行性疾病。