Ataxin-2 complex proteins in neurodegeneration.

项目总结/摘要 该R35提案建立在Javits R37和U 01 CREATE Bio奖支持的实验基础上， 长期以来，主要研究人员的小组从机械发现中取得了成功， 脊髓小脑共济失调和翻译到新的治疗方法，其中一个现在是在第一阶段的人体试验 （BIIB105）。该提案的统一主题是关注ATXN 2复合物中的RNA结合蛋白（RBP）， 指导RNA代谢的几个方面并且易于相分离的蛋白质， 聚合来基于我们最近在聚谷氨酰胺介导的神经变性和神经元变性方面的发现， staufen-1（STAU 1）过量，我们将应用多维度的方法来定义新的致病性 与自噬、未折叠蛋白反应（UPR）和RNA相互作用的机制 代谢和运输。我们假设这些反应虽然在短期内是补偿性的， 适应不良与持续的压力和RBP网络的反应导致进行性恶化， 自噬通量和凋亡信号的放大。我们已经建立了一套广泛的创新工具 和独特的模型在一个特殊的研究环境与建立当地和国家的合作者 多年来我们一直与他们共享资源。我们的方法是描述小脑 使用全基因组转录组学、蛋白质组学、形态学、生理学和生物学方法， 和行为技术。这些基础将使我们能够从细胞模型快速发展到动物模型 在体外和体内改变特定基因的剂量。我们现在将这种方法从浦肯野细胞扩展到 脊髓运动神经元使我们能够解决与遗传性和散发性疾病广泛相关的基本问题， 神经退行性疾病这些热点问题包括：细胞质RBP的作用，特别是ATXN 2 和STAU 1，响应营养，化学和突变蛋白质胁迫;它们的调节和相互作用， 相互作用，以及决定自噬通量和UPR的关键蛋白质;它们对神经元死亡的总体影响; 最后，它们有望成为小分子和RNA导向疗法的治疗靶点。R35 资金将使我们能够确定ATXN 2复合物中蛋白质在神经变性中的新功能， 定义不同神经退行性疾病的共同特征，不仅与 运动神经元疾病是一种遗传性疾病，但也与散发性共济失调和运动神经元疾病有关。我们建议的广泛范围 研究将使具有不同背景的科学家能够参与实验室和部门文化 包容性和多样性。凭借我们对动物模型和基因的可重复性的既定承诺， 针对性的方法，新的机械见解有希望翻译成开发新的 神经退行性疾病的治疗。

Project Summary/Abstract This R35 proposal builds on experiments supported by a Javits R37 and a U01 CREATE Bio award and the longstanding success of the principal investigator’s group leading from mechanistic discoveries in spinocerebellar ataxias and translation to novel treatments, one of which is now in a phase 1 human trial (BIIB105). The proposal’s unifying theme is a focus on RNA-binding proteins (RBPs) in the ATXN2-complex of proteins that direct several aspects of RNA metabolism and that are prone to phase-separation and aggregation. Building on our recent discoveries in polyglutamine-mediated neurodegeneration and neuronal staufen-1 (STAU1) overabundance, we will apply multi-dimensional approaches to define novel pathogenic mechanisms in their intersections with autophagy, the unfolded protein response (UPR), and with RNA metabolism and transport. We postulate that these responses, while compensatory in the short term, become maladaptive with sustained stress and that an RBP network response leads to progressive deterioration of autophagic flux and amplification of apoptotic signaling. We have established a broad suite of innovative tools and unique models in an exceptional research environment with established local and national collaborators with whom we have shared resources over many years. Our approach has been to characterize cerebellar degeneration at multiple time points using genome-wide transcriptomic, proteomic, morphologic, physiologic and behavioral techniques. These foundations will allow us to rapidly progress from cellular to animal models modifying dosage of specific genes in vitro and in vivo. We will now extend this approach from Purkinje cells to spinal motor neurons enabling us to address fundamental issues of broad relevance to inherited and sporadic neurodegenerative diseases. These topical issues include: the role of cytoplasmic RBPs, especially ATXN2 and STAU1, in response to nutrient, chemical and mutant protein stress; their regulation and interplay with each other, and key proteins determining autophagic flux and the UPR; their overall effect on neuronal death; and finally, their promise as therapeutic targets using small molecules and RNA-directed therapies. R35 funding will allow us to identify novel functions of proteins in the ATXN2-complex in neurodegeneration and define shared features across different neurodegenerative diseases with particular relevance not only to inherited, but also to sporadic forms of ataxia and motor neuron disease. The broad scope of our proposed studies will enable participation of scientists with diverse backgrounds in a laboratory and departmental culture of inclusivity and diversity. With our established commitment to reproducibility of animal models and gene targeting approaches, the novel mechanistic insights hold promise for translation into developing novel treatments for neurodegenerative disorders.