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

项目摘要 肌萎缩侧索硬化症（ALS）和额颞叶痴呆症（FTD）是相关的，通常是共病的， 没有可用的疾病改善疗法的神经退行性疾病。超过95%的ALS患者， 50%的FTD患者，受影响的神经元表现出胞质错误定位和TDP 43积聚 （反式反应元件DNA/RNA结合蛋白，43 kDa）。TARDBP基因突变， TDP 43也引起ALS和FTD的家族性形式，突出了TDP 43对这些疾病的整体贡献。 条件即便如此，疾病中TDP 43错误定位的潜在机制仍不清楚。 我们实验室最近的证据表明，TDP 43的错误定位可能是由于产生了 选择性剪接的缩短的TDP 43同种型，其从细胞核主动输出并倾向于 聚合来sTDP 43同种型在进化上也是保守的，但它们的调节和功能仍然存在 基本上未知。 该提议试图阐明sTDP 43同种型的不同功能及其对免疫调节的潜在贡献。 通过（i）定义sTDP 43的天然转录物和蛋白质相互作用物;和（ii）研究潜在的 sTDP 43在调节RNA稳定性中的作用。这样做，这些研究可能会揭示新的途径， TDP 43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的技能，将是 作为一名专注于分子生物学的独立研究者， 神经退化的基础

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.