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

项目总结 肌萎缩侧索硬化症(ALS)和额颞叶痴呆(FTD)是相关的，通常是并存的， 神经退行性疾病，没有可用的疾病修正疗法。在超过95%的ALS患者中，以及 50%的FTD患者，受累神经元出现胞浆错误定位和TDP43积聚 (反式反应元件DNA/RNA结合蛋白，43 kDa)。编码基因TARDBP的突变 TDP43也导致家族性ALS和FTD，突出了TDP43对这些疾病的整体贡献 条件。尽管如此，TDP43在疾病中错位的机制仍然不清楚。 我们实验室最近的证据表明，TDP43错误定位可能是由于产生了 另一种剪接的、缩短的(S)TDP43亚型，活跃地从核中输出并易于 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 根本不为人所知。 这一建议试图阐明sTDP43亚型的独特功能及其对 疾病通过(I)定义sTDP43的天然转录物和蛋白质相互作用；以及(Ii)研究潜在的 STDP43在调节RNA稳定性中的作用。在这样做的过程中，这些研究可能会揭示导致 TDP43错位和神经退行性变ALS和FTD。此外，它将使我能够开发出必要的 具备生物信息学、蛋白质组学、实验设计、数据分析和科学交流方面的技能 对我作为一名专注于分子研究的独立调查员的职业生涯的成功至关重要 神经退行性变的基础。

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.