Identifying the function of alternatively spliced TDP43 isoforms and contribution to disease

确定选择性剪接 TDP43 亚型的功能及其对疾病的影响

• 批准号: 10748166
• 负责人: Megan Dykstra
• 金额: $ 4.09万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748166
• 关键词: ALS patients; Adaptor Signaling Protein; Affect; Alternative Splicing; Amino Acids; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Binding; Bioinformatics; Biology; C-terminal; Cell Nucleus; Communication; Coupled; Cytoplasm; Cytoplasmic Granules; DNA; Data; Data Analyses; Disease; Event; Exhibits; Experimental Designs; Fluorescent in Situ Hybridization; Frontotemporal Dementia; G3BP1 gene; Genes; Glycine; Goals; Homeostasis; Immunoprecipitation; Individual; Induced pluripotent stem cell derived neurons; Introns; Investigation; Label; Laboratories; Length; Measures; Mediating; Messenger RNA; Metabolic; Methods; Molecular; Motor Neurons; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Nuclear; Nuclear Export; Nuclear Protein; Organelles; Pathogenesis; Pathology; Pathway interactions; Patients; Population; Production; Protein Isoforms; Proteins; Proteomics; RNA; RNA Binding; RNA Degradation; RNA Recognition Motif; RNA Splicing; RNA Stability; RNA-Binding Proteins; Regulation; Research Personnel; Response Elements; Ribonucleoproteins; Role; Services; Set protein; Testing; Toxic effect; Transcript; Translational Repression; Translations; Variant; Work; career; cell type; comorbidity; crosslink; frontotemporal lobar dementia amyotrophic lateral sclerosis; immunocytochemistry; insight; knock-down; mRNA Decay; mRNA Precursor; neuron loss; next generation; next generation sequencing; novel; novel therapeutic intervention; protein TDP-43; protein protein interaction; recruit; skills; stress granule; success; training opportunity; transcriptome sequencing

PROJECT SUMMARY Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are related, often comorbid, neurodegenerative disorders with no available disease-modifying therapies. In over 95% of ALS patients, and 50% of FTD patients, affected neurons exhibit cytoplasmic mislocalization and accumulation of TDP43 (transactive response element DNA/RNA binding protein, 43 kDa). Mutations in TARDBP, the gene encoding TDP43, also cause familial forms of ALS and FTD, highlighting an integral contribution of TDP43 to these conditions. Even so, the mechanisms underlying TDP43 mislocalization in disease remain unclear. Recent evidence from our laboratory suggests that TDP43 mislocalization may be due to the production of alternatively spliced, shortened (s)TDP43 isoforms that are actively exported from the nucleus and prone to aggregation. sTDP43 isoforms are also evolutionarily conserved, but their regulation and function remain fundamentally unknown. This proposal seeks to elucidate the distinct function of sTDP43 isoforms and their potential contribution to disease by (i) defining the native transcript and protein interactors for sTDP43; and (ii) investigating a potential role for sTDP43 in regulating RNA stability. In so doing, these studies may reveal new pathways responsible for TDP43 mislocalization and neurodegeneration ALS and FTD. Additionally, it will enable me to develop essential skills in bioinformatics, proteomics, experimental design, data analysis, and scientific communication that will be critical for my success in my intended career as an independent investigator focused on the molecular underpinnings of neurodegeneration.

项目摘要 肌萎缩性外侧硬化症（ALS）和额颞痴呆（FTD）相关，通常合并 神经退行性疾病，没有可用的疾病修改疗法。在超过95％的ALS患者中， 50％的FTD患者，受影响的神经元表现出细胞质错误和TDP43的积累 （交易反应元件DNA/RNA结合蛋白，43 kDa）。 TARDBP突变，基因编码 TDP43，也引起家族形式的ALS和FTD，突出了TDP43对这些的积分贡献 状况。即便如此，疾病中TDP43错误定位的基础机制仍不清楚。 我们实验室的最新证据表明，TDP43错误定位可能是由于生产 替代地剪接的，缩短的（S）TDP43同工型，这些同工型被积极从细胞核导出并容易出口到 聚合。 STDP43同工型也是进化保守的，但它们的调节和功能仍然保持 根本未知。 该建议旨在阐明STDP43同工型的独特功能及其对 通过（i）定义STDP43的天然转录本和蛋白质相互作用的疾病； （ii）研究潜力 STDP43在调节RNA稳定性中的作用。这样，这些研究可能揭示了负责的新途径 TDP43错误定位和神经变性ALS和FTD。此外，这将使我能够发展重要 具有生物信息学，蛋白质组学，实验设计，数据分析和科学沟通的技能 我作为我作为独立研究人员的意图职业至关重要的，专注于分子 神经退行性的基础。