RNA Dominant Mechanisms in ALS

ALS 中的 RNA 主导机制

• 批准号: 8541466
• 负责人: Peter K Todd
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-10-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8541466
• 关键词: Accounting; Affect; Age; American; Amyotrophic Lateral Sclerosis; Animal Model; Binding; CGG repeat; Cause of Death; Cell Culture Techniques; Cells; Cessation of life; Computer Simulation; Disease; Disease model; Drosophila genus; Exhibits; Eye; FXTAS; Frontotemporal Dementia; Future; Genetic; Glean; Goals; Grant; Health; Homeostasis; Human; In Vitro; Inherited; Introns; Lead; Longevity; Mediating; Methods; Modeling; Morbidity - disease rate; Motor Neuron Disease; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear; Nucleotides; Pathogenesis; Pathologic; Pathology; Patients; Process; Protein Binding; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; RNA-Protein Interaction; Research; Therapeutic; Tissues; Toxic effect; Translating; United States; Veterans; base; clinically relevant; disability; in vivo; mortality; motor neuron function; nervous system disorder; neurotoxicity; novel; prevent; protein TDP-43; protein function; research study; small molecule; therapeutic development

DESCRIPTION (provided by applicant): Alterations in RNA homeostasis and RNA binding protein functions underlie critical aspects of pathogenesis in neurodegeneration. For RNA dominant disorders caused by nucleotide repeats, the effects of RNA are direct, with the repeats themselves eliciting toxicity via the sequestration of certain proteins. For other neurodegenerative conditions, including amyotrophic lateral sclerosis (ALS), RNA binding proteins such as TDP-43 form neuronal aggregates that serve as pathologic hallmarks of disease. Recently, a novel intronic GGGGCC nucleotide repeat expansion (C9ALS) was identified as the most common known cause of ALS. C9ALS is associated with TDP-43 pathology but also exhibits accumulation of repeat RNA into nuclear foci. We propose a set of studies that take advantage of our expertise in RNA dominant disorders to interrogate how the GGGGCC repeat elicits disease in C9ALS. We hypothesize that GGGGCC repeats are inherently toxic as RNA, and that they elicit toxicity by sequestering specific RNA binding proteins. The immediate goals of this study are to generate and characterize a drosophila model of C9ALS and to identify proteins which interact with the GGGGCC repeat in vitro and in vivo. Research Plan/Methods: To better understand C9ALS pathogenesis, we will first generate an GGGGCC repeat model in Drosophila that allows us to track both the splicing and localization of the intron within expressing tissues. We will determine if GGGGCC RNA repeats confer neurodegeneration and affect lifespan and motor neuron function. We will then take a candidate approach to identify genetic suppressors of GGGGCC repeat toxicity. In a parallel set of studies, we will use in vitro and cell culture based techniqes to empirically identify proteins that bind to the GGGGCC repeat. We will then validate that these candidates colocalize with RNA inclusions in Drosophila and patient cells. Impact/Clinical Relevance: All forms of ALS and FXTAS are currently untreatable and impart significant morbidity and mortality upon Veterans. The C9ALS repeat discovery provides a unique opportunity to rapidly translate recent advances in RNA dominant diseases to this new disorder. The long term goal of this project is to identify small molecules that block interactions between GGGGCC repeat RNA with its cognate RNA binding proteins. Such compounds will then be evaluated for their ability to prevent GGGGCC RNA toxicity in animal models and eventually utilized to treat patients with C9ALS. Taken together, our approach is novel, feasible, and of high potential impact to patients with ALS and related GGGGCC associated neurological disorders.

描述（由申请人提供）： RNA稳态和RNA结合蛋白功能的改变是神经退行性中发病机理的关键方面的基础。对于由核苷酸重复序列引起的RNA显性疾病，RNA的作用是直接的，重复序列本身会通过隔离引起毒性 某些蛋白质。对于其他神经退行性疾病，包括肌萎缩性侧性硬化症（ALS），RNA结合蛋白（例如TDP-43）形成了神经元聚集体，这些神经元聚集体是病理学标志的疾病。 最近，一种新型的内含子GGGGCC核苷酸重复扩张（C9AL）被确定为最常见的ALS原因。 C9AL与TDP-43病理学有关，但也表现出重复RNA的积累到核灶中。我们提出了一系列研究，以利用我们在RNA显性疾病中的专业知识，以询问GGGGCC如何重复在C9ALS中引起疾病​​。我们假设GGGGCC重复固有地是RNA的毒性，并且它们通过隔离特定的RNA结合蛋白引起毒性。这项研究的直接目标是生成和表征C9AL的果蝇模型，并鉴定与GGGGCC在体外和体内重复相互作用的蛋白质。研究计划/方法：为了更好地了解C9als的发病机理，我们将首先在果蝇中生成GGGGCC重复模型，使我们能够跟踪表达组织中内含子的剪接和定位。我们将确定 如果GGGGCC RNA重复赋予神经变性并影响寿命和运动神经元功能。然后，我们将采用一种候选方法来鉴定GGGGCC重复毒性的遗传抑制剂。 在一系列平行的研究中，我们将使用基于细胞和细胞培养的技术来鉴定与GGGGCC重复结合的蛋白质。然后，我们将验证这些候选者与果蝇和患者细胞中的RNA夹杂物共定位。影响/临床相关性：所有形式的ALS和FXTA目前都无法治疗，并且对退伍军人施加了明显的发病率和死亡率。 C9als重复发现提供了一个独特的机会，可以快速将RNA主要疾病的最新进展转化为这种新疾病。该项目的长期目标是确定与GGGGCC重复RNA与其同源RNA结合蛋白之间相互作用的小分子。然后，将评估此类化合物的预防动物模型中GGGGCC RNA毒性的能力，并最终用于治疗C9ALS患者。综上所述，我们的方法是新颖，可行的，并且对ALS和相关GGGGCC相关的神经系统疾病的患者具有很高的潜在影响。