Cellular and Molecular Mechanisms of FUS-related ALS/FTD

FUS相关ALS/FTD的细胞和分子机制

基本信息

批准号：
9763021
负责人：
Udai B Pandey
金额：
$ 38.41万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-15 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9763021
关键词：
Amyotrophic Lateral Sclerosis Animal Model Animals Award Axon Behavioral Assay Biochemical Biogenesis Biological Assay Brain CAG repeat Cell Culture Techniques Cell model Cell physiology Complex Cytoplasm Cytoplasmic Granules Defect Disease Drosophila genus Ectopic Expression Frontotemporal Dementia Functional disorder Genes Genetic Genetic Screening Genetic Transcription Goals Homologous Gene Human Light Link Longevity Lower Organism Mammalian Cell Mediating Metabolism MicroRNAs Modeling Molecular Motor Neurons Mutate Mutation Myotonic Dystrophy Nerve Degeneration Neurodegenerative Disorders Neuromuscular Junction Neurons Nuclear Pathogenesis Pathogenicity Pathologic Pathology Pathway interactions Patients Physiological Proteins RNA RNA Interference RNA Splicing RNA-Binding Protein FUS RNA-Binding Proteins Small Nuclear RNA Spinal Muscular Atrophy Structure Therapeutic Intervention Toxic effect Translations Ubiquitin Vertebrates Work cellular pathology cofactor disease-causing mutation fly frontotemporal degeneration frontotemporal lobar dementia-amyotrophic lateral sclerosis human disease in vivo induced pluripotent stem cell knock-down molecular pathology motor neuron degeneration mutant novel patient subsets protein TDP-43 recruit stress granule trafficking transcriptome transcriptome sequencing transcriptomics

项目摘要

ALS is a late-onset progressive neurodegenerative disease caused by degeneration of motor neurons, and a disease hallmark is the accumulation of ubiquitin-positive aggregates in neuronal cytoplasm. FUS was identified as genes mutated in both familial and sporadic forms of ALS. In fact, a subset of patients with frontotemporal dementia (FTD) show FUS pathology. FUS, similar to TDP-43, is an RNA binding protein implicated in multiple aspects of RNA metabolism, including splicing, trafficking, and translation. The precise mechanisms of mutated FUS in ALS pathogenesis are not known. To understand the molecular mechanisms of FUS-mediated neurodegeneration, we developed cellular (mammalian primary neuronal and patient-derived iPSC motor neuron) and Drosophila models that recapitulate key features of human disease including cytoplasmic mislocalization, neuromuscular junction defects, locomotor dysfunctions, reduced life span, perturbed stress granule dynamics and toxicity. We discovered muscleblind and drosha as unexpected and novel modifiers of mutant FUS toxicity. MBNL proteins, highly conserved from lower organisms to vertebrates, have been implicated in many neurodegenerative disorders, such as myotonic dystrophy and CAG repeat diseases. The long-term goal is to identify modifiers of FUS toxicity and understand their molecular mechanisms using mammalian cell culture and Drosophila models. The objective of our current application is to determine how muscleblind and drosha modulate FUS-mediated toxicity in Drosophila and FUS iPSC motor neurons. We hypothesize that muscleblind and drosha regulate RNA splicing, SG dynamics and miRNA biogenesis that is perturbed by pathogenic mutations in FUS. We will examine the impact of muscleblind and drosha on cellular and molecular pathologies in FUS-associated neurodegeneration. We expect to dissect the molecular pathways that could be exploited for developing therapeutic interventions for ALS/FTD patients.

ALS是由运动变性引起的晚期发作的进行性神经退行性疾病神经元和疾病标志是泛素阳性聚集体的积累神经元细胞质。 FUS被鉴定为在家族性和零星形式中突变的基因 ALS。实际上，额颞痴呆（FTD）的一部分患者表现出FUS病理。类似于TDP-43的FUS是一种与RNA多个方面有关的RNA结合蛋白代谢，包括剪接，贩运和翻译。突变的精确机制 ALS发病机理中的FUS尚不清楚。为了了解FUS介导的神经变性的分子机制，我们发展了细胞（哺乳动物原发性神经元和患者衍生的IPSC运动神经元）和果蝇概括人类疾病关键特征的模型，包括细胞质错误定位，神经肌肉连接缺陷，运动功能障碍，寿命降低，干扰应力颗粒动力学和毒性。我们发现Muscleblind和Drosha是出乎意料而新颖的突变FUS毒性的修饰符。 MBNL蛋白，从低生物体高度保守脊椎动物已与许多神经退行性疾病有关营养不良和CAG重复疾病。长期目标是确定FUS毒性的修饰符并使用哺乳动物细胞培养和果蝇了解它们的分子机制型号。我们当前应用的目的是确定肌肉盲和drosha如何调节果蝇和FUS IPSC运动神经元的FUS介导的毒性。我们假设这种肌肉蓝线和Drosha调节RNA剪接，SG动力学和miRNA生物发生， FUS中的致病突变受到干扰。我们将研究肌肉盲和 DROSHA在FUS相关神经退行性中的细胞和分子病理学上。我们期望剖析可以利用用于发展治疗的分子途径 ALS/FTD患者的干预措施。