mRNA Targets for TDP-43 and FUS/TLS: Identifying Key RNA-Processing Errors in ALS

TDP-43 和 FUS/TLS 的 mRNA 靶标：识别 ALS 中的关键 RNA 处理错误

• 批准号: 8263409
• 负责人: Magdalini Polymenidou
• 金额: $ 9.1万
• 依托单位: LUDWIG INSTITUTE FOR CANCER RES LTD
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8263409
• 关键词: Adult; Affect; Alternative Splicing; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Brain; Cell Culture Techniques; Cell Nucleus; Cessation of life; Complementary DNA; Coupled; Cytoplasmic Inclusion; Diagnosis; Disease; Employee Strikes; Event; Fibroblasts; Frontotemporal Dementia; Genetic Transcription; Human; Immunoprecipitation; Interruption; Intervention; Link; Messenger RNA; Metabolism; Methodology; Motor Neurons; Mus; Muscle Weakness; Mutation; Nerve Degeneration; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neurons; Paralysed; Parkinson Disease; Pathogenesis; Patients; Pattern; Physiological; Pluripotent Stem Cells; Proteins; Public Health; RNA; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regulation; Role; Technology; Testing; Therapeutic Intervention; Transcript; Transgenic Mice; Work; crosslink; disease-causing mutation; effective therapy; efficacy testing; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; induced pluripotent stem cell; loss of function; mouse model; mutant; premature; protein TDP-43; public health relevance; stem

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder in which premature loss of upper and lower motor neurons leads to fatal paralysis with a typical disease course of one to five years. Starting from 2006 with the identification of TDP-43 as the major protein component of the ubiquitinated inclusions occurring in affected neurons of ALS and Frontotemporal Lobar Dementia (FTLD) patients, there has been what is widely recognized to be a paradigm-shifting breakthrough in our understanding of neurodegeneration. Indeed, it has now been established that TDP-43, an RNA-binding protein that is normally localized mainly in the nucleus, forms cytoplasmic aggregates, which alter its subcellular localization in a variety of different neurodegenerative conditions, including ALS, FTLD, Alzheimer's and Parkinson's disease. In addition, identification of ALS-causing mutations not only in TDP-43, but more recently, also in another RNA-binding protein, namely FUS/TLS, confirmed the primary role of these proteins in the pathogenesis of ALS. Furthermore, these discoveries solidified the message of a key contribution of RNA-processing alterations to mechanisms of neurodegeneration. I plan to define a set of disease-relevant TDP-43- and/or FUS/TLS-dependent RNA-processing alterations. I propose to utilize existing transgenic mouse lines along with newly developed methodologies including high-throughput sequencing and induced pluripotent stem (iPS) cells to 1) determine the role of TDP-43 and FUS/TLS in the regulation of RNA metabolism in the normal central nervous system in mice, 2) identify RNA-processing alterations caused by expression of TDP-43 and FUS/TLS carrying disease-causing mutations in transgenic mice, 3) test the RNA-processing changes found in mice, in human motor neurons produced from iPS cells, and 4) identify the mechanisms underlying the observed disease-related RNA-processing alterations and test the efficacy of blocking compounds on the defined mechanisms in cell culture and mouse models. PUBLIC HEALTH RELEVANCE: Statement of Relevance to Public Health Amyotrophic Lateral Sclerosis, also called 'Lou Gehrig's disease', is a devastating disease causing muscle weakness progressing to paralysis and death within 1-5 years from diagnosis. There are no effective treatments for this disease that affects 1 in approximately 55,000 people in USA every year. The proposed work will contribute to our understanding of the chain of events leading to this disease and to uncover targets for therapeutic interventions.

描述（由申请人提供）： 肌萎缩侧索硬化症（ALS）是一种成年发病的神经退行性疾病，其中上和下运动神经元的过早丧失导致致命的瘫痪，典型的病程为1至5年。从2006年开始，随着TDP-43被鉴定为ALS和额颞叶痴呆（FTLD）患者受影响神经元中出现的遍在蛋白化内含物的主要蛋白成分，人们普遍认为这是一个范式转变突破。我们对神经退行性疾病的理解。事实上，现已确定TDP-43（一种通常主要定位于细胞核中的RNA结合蛋白）形成细胞质聚集体，这改变了其在多种不同神经退行性病症（包括ALS、FTLD、阿尔茨海默病和帕金森病）中的亚细胞定位。此外，不仅在TDP-43中，而且最近还在另一种RNA结合蛋白（即FUS/TLS）中鉴定了ALS引起的突变，证实了这些蛋白在ALS发病机制中的主要作用。此外，这些发现巩固了RNA加工改变对神经变性机制的关键贡献的信息。我计划定义一组疾病相关的TDP-43和/或FUS/TLS依赖的RNA加工改变。我建议利用现有的转基因小鼠品系沿着新开发的方法，包括高通量测序和诱导多能干细胞（iPS），以1）确定TDP-43和FUS/TLS在小鼠正常中枢神经系统中调节RNA代谢的作用，2）在转基因小鼠中鉴定由携带致病突变的TDP-43和FUS/TLS的表达引起的RNA加工改变，3）测试在小鼠中、在由iPS细胞产生的人运动神经元中发现的RNA加工变化，以及4）鉴定所观察到的疾病相关的RNA加工改变的潜在机制，并测试阻断化合物对细胞培养物和小鼠模型中所定义的机制的功效。 公共卫生关系：肌萎缩性侧索硬化症（Amyotrophic Lateral Sclerosis，又称“Lou Gehrig's disease”）是一种破坏性疾病，在确诊后1-5年内会导致肌肉无力、瘫痪和死亡。没有有效的治疗方法，这种疾病每年影响美国约55，000人中的1人。拟议的工作将有助于我们了解导致这种疾病的事件链，并发现治疗干预的目标。