Elucidating the role of RNA oxidation on amyotrophic lateral sclerosis onset and progression

阐明 RNA 氧化对肌萎​​缩侧索硬化症发病和进展的作用

• 批准号: 10371642
• 负责人: Isaac Alexander Chaim
• 金额: $ 12.45万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371642
• 关键词: Aging; Amyotrophic Lateral Sclerosis; Amyotrophic Lateral Sclerosis Pathway; Biotechnology; Cells; Computing Methodologies; DNA; DNA Repair; Disease; Disease Progression; Environment; Faculty; Foundations; Genes; Genetic Transcription; In Vitro; Individual; Institutes; Intervention; Lead; Link; Location; Mentors; Methods; Modeling; Molecular; Motor Neurons; Mutation; Nerve Degeneration; Neurons; Organoids; Oxidative Stress; Oxides; Pathogenesis; Patients; Positioning Attribute; Proteins; RNA; RNA Processing; RNA immunoprecipitation sequencing; RNA-Binding Proteins; Reactive Oxygen Species; Regulation; Research; Research Institute; Research Personnel; Resolution; Role; Specimen; Spinal; Spinal Cord; Testing; Therapeutic Intervention; Time; Tissues; Training; Transcript; Translations; Work; activation-induced cytidine deaminase; cell injury; cell type; diagnostic strategy; familial amyotrophic lateral sclerosis; high risk; induced pluripotent stem cell; insight; motor neuron degeneration; mouse model; mutant; neuron loss; novel; oxidation; programs; response; sporadic amyotrophic lateral sclerosis; transcriptomics

PROJECT SUMMARY A growing body of evidence implicates oxidative stress in Amyotrophic Lateral Sclerosis (ALS). Nevertheless, it is currently unknow if it is a cause, a by-product or a consequence of disease. The effects of oxidative stress on cellular damage caused by reactive oxygen species is usually attributed to modifying proteins and DNA. However, RNA oxidation occurs ten times more often than DNA oxidation. Importantly, high levels of oxidized RNA are detected in relevant neuronal tissues of patients with ALS while, mouse models of ALS show increased RNA oxidation in motor neurons of the spinal cord at an early pre-symptomatic stage. What remains lacking, however, is an understanding of the functional relationship between RNA oxidation and ALS onset and progression. Thus, there is a critical need to identify which motor neuron transcripts are oxidized in early, pre- symptomatic stages of ALS and how this dysregulation contributes to neuronal death and other molecular hallmarks of ALS. I believe that there are many, yet to be discovered, RNA Binding Proteins (RBPs) that are crucial for controlling the fate of oxidized RNAs. I hypothesize that RNA oxidation drives motor neuron degeneration in ALS by dysregulating proper RNA processing by RBPs. I will test this hypothesis by (1) elucidating, in iPSC-derived motor neurons, the RNA targets and the consequences of depletion of known RBPs that interact with oxidized RNAs and identify novel ones; (2) identifying and comparing oxidized RNAs in iPSC- derived motor neuron models; and (3) investigating, at a single cell resolution, the effects of RNA oxidation on transcription, translation and RBP-RNA interactions in iPSC-derived spinal organoid ALS models. If successful, this project will generate the foundational methods and insights to enable early, pre-symptomatic-stage diagnostic approaches and interventions to reduce RNA oxidation levels in high-risk individuals. My background in DNA damage and repair and single cell transcriptomics together with the Yeo lab’s expertise on RNA processing and neurodegeneration make me an ideal candidate to accomplish the research proposed above. These three aims will serve as a basis for my independent academic position generating the foundational methods and insights to study the effects of RNA damage. The Yeo lab at UCSD is an ideal environment to perform this research and complete my training towards pursuit of an independent academic faculty position, as it has consistently been a leader in developing both experimental and computational methods to characterize RNA processing and RBP regulation. Additionally, the location of the Yeo lab proximal to outstanding researchers at UCSD, the Salk Institute, and other research institutes and biotechnology companies in La Jolla will provide ample opportunities for mentored training in performing research and developing an independent research program.

项目摘要 越来越多的证据表明氧化应激与肌萎缩侧索硬化症（ALS）有关。但它 目前尚不清楚它是疾病原因、副产品还是后果。氧化应激对 由活性氧物质引起的细胞损伤通常归因于修饰蛋白质和DNA。 然而，RNA氧化发生的频率是DNA氧化的十倍。重要的是，高水平的氧化 在ALS患者的相关神经元组织中检测到RNA，而ALS的小鼠模型显示增加的 早期症状前阶段脊髓运动神经元的RNA氧化。剩下的不足， 然而，理解RNA氧化和ALS发作之间的功能关系， 进展因此，迫切需要确定哪些运动神经元转录本在早期、前-晚期被氧化。 ALS的症状阶段以及这种失调如何导致神经元死亡和其他分子 ALS的标志我相信有许多尚未被发现的RNA结合蛋白（RBP）， 对于控制氧化RNA的命运至关重要。我假设RNA氧化驱动运动神经元 通过RBP对适当RNA加工的失调而导致ALS变性。我将通过（1）来检验这个假设。 在iPSC衍生的运动神经元中阐明RNA靶点和已知RBP耗尽的后果 与氧化RNA相互作用并鉴定新的RNA;（2）鉴定和比较iPSC中的氧化RNA- 衍生的运动神经元模型;和（3）研究，在单细胞分辨率，RNA氧化的影响， 在iPSC衍生的脊髓类器官ALS模型中的转录、翻译和RBP-RNA相互作用。如果成功， 该项目将产生基本的方法和见解，使早期，前症状阶段， 诊断方法和干预措施，以降低高风险个体的RNA氧化水平。 我在DNA损伤和修复以及单细胞转录组学方面的背景以及Yeo实验室的专业知识 RNA加工和神经退行性变的研究使我成为完成这项研究的理想人选 以上这三个目标将作为我独立学术立场的基础， 研究RNA损伤影响的方法和见解。加州大学圣地亚哥分校的Yeo实验室是一个理想的环境， 进行这项研究，并完成我的培训，追求一个独立的学术教师的立场，作为 它一直是开发实验和计算方法的领导者， RNA加工和RBP调节。此外，杨实验室的位置接近突出的 加州大学圣地亚哥分校、索尔克研究所和其他研究机构以及拉霍亚的生物技术公司的研究人员 将提供充足的机会，在执行研究和开发一个独立的指导培训 研究计划。