Using RNA signatures for therapy development in neurodegeneration due to C9orf72 expansions

使用 RNA 特征开发 C9orf72 扩展引起的神经退行性疾病的治疗方法

基本信息

批准号：
8817335
负责人：
Clotilde Lagier-Tourenne
金额：
$ 28.02万
依托单位：
LUDWIG INSTITUTE FOR CANCER RES LTD
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8817335
关键词：
Affect Amyotrophic Lateral Sclerosis Antisense Oligonucleotides Binding C9ORF72 Cells Chemicals Clinical Trials Collaborations Consensus Dipeptides Disease Employee Strikes Engineering FDA approved Familial Amyotrophic Lateral Sclerosis Fibroblasts Frontotemporal Dementia Functional RNA Genes Genetic Genetic Transcription Genome Genomics High-Throughput Nucleotide Sequencing Human In Vitro Individual Link Mediating Messenger RNA Methods Molecular Profiling Motor Cortex Motor Neurons Nerve Degeneration Neurodegenerative Disorders Neurons Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Play Preclinical Drug Evaluation Process Production Property Proteins RNA RNA Processing RNA Splicing RNA-Binding Proteins Reporting Role Seminal Spinal System Testing Therapeutic Toxic effect Transcript Translating Translations Work cost effective cytotoxic cytotoxicity design drug efficacy gain of function high throughput screening induced pluripotent stem cell innovation loss of function novel therapeutics public health relevance screening small molecule therapeutic development therapy development

项目摘要

DESCRIPTION (provided by applicant): RNA processing alterations are increasingly recognized to play a crucial role in the pathogenesis of a wide range of diseases including two devastating neurodegenerative conditions, amyotrophic lateral sclerosis (ALS) and frontal temporal dementia (FTD). The seminal discovery in 2011 of a hexanucleotide expansion in the C9orf72 gene as the most common cause of familial ALS and FTD significantly changed our perspective of these neurodegenerative diseases. The pathogenic mechanisms of this expansion are not yet understood. However recent lines of evidence, including from my own work, strongly support a gain of toxic property by accumulation of repeat-containing RNAs that are bidirectionally transcribed from the C9orf72 locus. Independent reports of RNA changes in fibroblasts, neurons derived from iPS cells and motor cortex from C9orf72 patients have offered evidence that RNA processing misregulation accompanies C9orf72 disease. However, it is still unresolved whether these RNA alterations are induced by the functional disruption of one or more RNA binding proteins sequestered into RNA foci or induced by another cytotoxic mechanism such as the striking accumulation of dipeptide repeat proteins translated from expanded RNAs that was recently reported in C9orf72 patients. In this project, I propose to define a set of alterations in mRNA processing that delineate a disease-dependent signature and that will serve as functional readouts to distinguish among current hypotheses for disease mechanism underlying C9orf72 ALS/FTD. By using genomic approaches optimized for precise identification of splicing and expression changes and human neurons specifically engineered to individually explore toxic mechanisms currently proposed, my group will determine which factor(s) drive the emergence of RNA processing alterations in C9orf72 disease. In particular, we will test the hypothesis that RNA alterations are induced either by sense or antisense repeat- containing RNAs. We will also use a functional screen to determine whether C9orf72-related RNA alterations are driven by reduced activity of one or more RNA binding proteins, including the proteins recently found to interact with the C9orf72 expanded repeats. Finally, my team will use the C9orf72 molecular signature to screen therapeutic compounds. Indeed, we and others have already established that antisense oligonucleotides (ASOs) targeting C9orf72 transcripts efficiently reduce pathological RNA foci in patient cells. I now propose to determine whether degradation of expanded RNAs transcribed from either the sense or antisense directions (or both), is necessary to restore RNA processing alterations linked to C9orf72 disease, an approach that may provide crucial information for the design of a clinical trial using ASOs in C9orf72 ALS/FTD patients. Disease-related RNA signatures will also serve as readouts to screen new therapeutic compounds in neurons directly derived from patient fibroblasts. Indeed, instead of using a single target RNA to determine drug efficacy, we will use quantitative analysis of a large panel of genes perturbed by C9orf72 expansions to identify small molecules that can intervene with disease-linked pathways to restore levels of the most affected RNAs.

描述（由申请人提供）：越来越多地认识到RNA加工改变在多种疾病的发病机理中起着至关重要的作用，包括两种毁灭性神经退行性疾病，肌萎缩性侧面硬化症（ALS）和额叶颞痴呆（FTD）。 2011年，C9ORF72基因中的六核苷酸扩展作为家族性ALS和FTD的最常见原因的开创性发现显着改变了我们对这些神经退行性疾病的看法。这种扩展的致病机制尚不清楚。然而，最近的证据（包括我自己的工作）是通过从C9ORF72基因座进行双向转录的重复含RNA来强烈支持有毒性能的。对成纤维细胞的RNA变化，IPS细胞衍生的神经元和C9ORF72患者的运动皮层的独立报道提供了证据，表明C9orf72疾病伴随着RNA处理不调节。然而，仍然尚未解决这些RNA改变是否是由于一种或多种RNA结合蛋白的功能破坏被隔离为RNA焦点的功能破坏还是由另一种细胞毒性机制诱导的诱导，例如从C9ORF72患者最近在C9ORF72患者中报告的二肽重复蛋白的震中积累。在这个项目中，我建议定义描述疾病依赖性特征的mRNA加工中的一系列变化，并将作为功能读数，以区分C9orf72 ALS/FTD的疾病机制的当前假设。通过使用优化的基因组方法来精确鉴定剪接和表达变化以及专门探索目前提出的有毒机制的人类神经元，我的组将确定哪些因素可以推动C9ORF72疾病中RNA处理改变的出现。特别是，我们将检验以下假设：RNA改变是通过含有含义的RNA诱导的。我们还将使用功能性屏幕来确定C9ORF72相关的RNA改变是否由一种或多种RNA结合蛋白的活性降低，包括最近发现与C9ORF72相互作用的蛋白。最后，我的团队将使用C9ORF72分子签名来筛选治疗化合物。的确，我们和其他人已经确定，针对C9orf72转录本的反义寡核苷酸（ASO）有效地降低了患者细胞中的病理RNA灶。我现在建议确定从感官或反义方向转录的扩展的RNA的降解（或两者兼而有之）对于恢复与C9ORF72疾病相关的RNA处理改变是必要的，这种方法可能为在C9ORF72 ALS/FTD患者中使用ASOS使用ASO的临床试验提供至关重要的信息。与疾病相关的RNA特征还将作为读取直接源自患者成纤维细胞的神经元中新的治疗化合物的读数。的确，我们将不用使用单个靶RNA来确定药物疗效，而是对C9ORF72扩张扰动的大型基因的定量分析来鉴定可以干预疾病连接途径的小分子，以恢复受影响最大的RNA的水平。