RNA decay in amyotrophic lateral sclerosis and frontotemporal lobar degeneration

摘要 维持RNA动态平衡涉及到RNA合成和周转之间的动态平衡。 这种平衡对神经元等转录活跃的细胞至关重要，对任何个体都是破坏。 成分可导致RNA错误处理和细胞死亡。我们的初步证据表明，缺陷 在RNA衰变中，代表了一种导致神经元功能障碍的基本且迄今未被认识的机制 神经退行性疾病肌萎缩侧索硬化症(ALS)和额颞叶的死亡 变性(FTLD)。我们提出这一建议的目的是阐明rna衰变在发病机制中的作用。 ALS和FTLD的研究，并研究一种影响RNA衰退的独特和有前途的治疗策略。基座 根据初步研究结果，我们认为在ALS和最常见的FTLD亚型中， 以神经元沉积RNA结合蛋白TDP43为特征，RNA动态平衡被一种 TDP43和RNA解旋酶UPF1之间的不适当相互作用，最终导致 神经退行性变。我们将通过以下方式测试这一模型：1)确定TDP43-UPF1相互作用对 神经元存活和RNA衰变，II)评估RNA衰变是否充分和/或必要 病理TDP43在神经元中的沉积，以及III)评估UPF1的表达是否改善了神经元 通过恢复TDP43和RNA动态平衡存活。我们将通过一系列措施来实现这些目标 ALS和FTLD患者来源的人类神经元的纵向单细胞显微镜，CRISPR/Cas9 基因组编辑和脉冲标记RNA的高通量测序。提出了多方面的方法 这里承诺揭示控制健康细胞和受疾病影响的神经元存活的关键途径 ALS和FTLD，并将使我们离实现开发有效的 治疗这些和其他坚持不懈的进行性神经退行性疾病。

Abstract Maintenance of RNA homeostasis involves a dynamic balance between RNA synthesis and turnover. This balance is critical for transcriptionally active cells such as neurons, as disruptions to any individual component can lead to RNA misprocessing and cell death. Our preliminary evidence indicates that deficiencies in RNA decay represent a fundamental and heretofore unrecognized mechanism driving neuronal dysfunction and death in the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Our goals with this proposal are to elucidate the role of RNA decay in the pathogenesis of ALS and FTLD, and investigate a unique and promising therapeutic strategy affecting RNA decay. Based upon the results of initial studies, we propose that in ALS and the most common subtype of FTLD, characterized by neuronal deposits of the RNA binding protein TDP43, RNA homeostasis is disrupted by an inappropriate interaction between TDP43 and the RNA helicase UPF1, ultimately resulting in neurodegeneration. We will test this model by i) determining the impact of the TDP43-UPF1 interaction on neuronal survival and RNA decay, ii) evaluating if deficient RNA decay is sufficient and/or necessary for pathologic TDP43 deposition in neurons, and iii) assessing whether UPF1 expression improves neuronal survival by restoring TDP43 and RNA homeostasis. We will pursue these aims using a combination of longitudinal single-cell microscopy of human neurons derived from ALS and FTLD patients, CRISPR/Cas9 genome editing and high-throughput sequencing of pulse-labeled RNA. The multifaceted approach proposed here promises to uncover key pathways controlling neuronal survival in healthy cells and in those affected by ALS and FTLD, and will bring us one step closer to achieving our long-term goal of developing an effective treatment for these and other relentlessly progressive neurodegenerative disorders.