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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆 (FTD) 是相关的，并且通常是共存的，没有可用的疾病缓解疗法的神经退行性疾病。超过 95% 的 ALS 患者，以及 50% 的 FTD 患者受影响的神经元表现出细胞质错误定位和 TDP43 积累（反式反应元件 DNA/RNA 结合蛋白，43 kDa）。 TARDBP（编码基因）突变 TDP43 也会导致家族性 ALS 和 FTD，这凸显了 TDP43 对这些疾病的整体贡献状况。即便如此，TDP43 在疾病中错误定位的机制仍不清楚。我们实验室的最新证据表明 TDP43 错误定位可能是由于产生选择性剪接、缩短的 (s)TDP43 同工型，从细胞核主动输出，易于发生聚合。 sTDP43 亚型在进化上也是保守的，但它们的调节和功能仍然存在根本不知道。该提案旨在阐明 sTDP43 同工型的独特功能及其对通过 (i) 定义 sTDP43 的天然转录物和蛋白质相互作用因子来识别疾病； (ii) 调查潜在的 sTDP43 在调节 RNA 稳定性中的作用。通过这样做，这些研究可能会揭示负责的新途径 TDP43 错误定位和神经变性 ALS 和 FTD。此外，它将使我能够开发必要的生物信息学、蛋白质组学、实验设计、数据分析和科学交流方面的技能作为一名专注于分子生物学的独立研究者，这对我在职业生涯中取得成功至关重要神经退行性变的基础。