权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanistic studies of ALS-causative mutations and RNP-focussed drug discovery using in vitro reconstitution of RNP complexes

使用 RNP 复合物的体外重建对 ALS 致病突变和以 RNP 为重点的药物发现进行机制研究

基本信息

批准号：
MR/W028522/1
负责人：
Tatyana Shelkovnikova
金额：
$ 47.74万
依托单位：
University of Sheffield
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2022
资助国家：
英国
起止时间：
2022 至无数据
项目状态：
未结题

来源：
https://gtr.ukri.org/projects?ref=MR%2FW028522%2F1
关键词：
Mechanistic studies ALS causative mutations

项目摘要

Amyotrophic lateral sclerosis (ALS), the most common form of motor neuron disease (also known as Lou Gehrig disease), is a fatal disease of the nervous system. ALS can be inherited or have no known genetic cause, yet all patients will develop a very similar condition. In all patents, nervous cells called motor neurons, mainly located in the spinal cord, become affected and eventually die. This leads to loss of support for muscles, causing their wasting, weakness and paralysis. Two drugs currently approved for ALS extend the lifespan by a few months and are not able to cure patients. Hence, there is a clear need for new drugs to treat ALS. ALS is characterised by abnormal metabolism of the molecular complexes in neurons composed of proteins and ribonucleic acids (RNAs), called ribonucleoprotein (RNP) complexes. Mutant, abnormally localised, or otherwise dysregulated proteins gain abnormal RNA-binding properties and structurally perturb RNP complexes leading to ALS pathology. Similarly, disease-relevant RNAs can establish unlicensed RNA-protein interactions with profound consequences for cellular RNA metabolism. We found that a specific RNA called NEAT1_2 that induces formation of RNP complexes known as paraspeckles might be an important player in ALS pathogenesis. Although absent from the neurons of healthy individuals, this RNA becomes accumulated in ALS motor neurons. In addition, many proteins that regulate NEAT1_2/paraspeckles are affected by ALS mutations. We believe it has a protective effect and helps neurons survive. However currently no NEAT1_2 targeted drugs are available.Traditional drug discovery efforts have focussed on the protein but it has recently become possible to identify drugs which act on RNA and its complexes with proteins. More specifically, drugs which affect RNPs are currently being evaluated in patients with the fatal childhood disease spinal muscular atrophy (SMA). Recently, we have developed a system that will allow us assembling complexes similar in composition to NEAT1_2/paraspeckles, by attaching a portion of NEAT1_2 RNA to micro-beads in a multi-well test plate. This system can be used for rapid analysis of protein binding to these complexes using an ultrafast microscopic imaging analysis. In the current proposal, we aim to better understand the link between NEAT1_2/paraspeckles and ALS, and find novel small molecules that can modulate these complexes with a potential to be used in ALS research and drug discovery. Further, we will modify this system to make it suitable for finding small molecules for any ALS-relevant RNA molecule using a major ALS-causative RNA produced from C9ORF72 gene. Our teams have significant background in RNA biology, ALS pathobiology and drug discovery as well as all the relevant laboratory tools. Therefore we are uniquely placed to use the above state-of-the-art approach we developed, to translate recent breakthroughs in our understanding of ALS pathophysiology into therapies development and ultimately, patient impact.

肌萎缩侧索硬化症（ALS）是运动神经元疾病（也称为卢伽雷病）最常见的形式，是一种致命的神经系统疾病。ALS可以遗传或没有已知的遗传原因，但所有患者都会发展出非常相似的病症。在所有患者中，主要位于脊髓中的称为运动神经元的神经细胞受到影响并最终死亡。这会导致失去对肌肉的支持，导致他们的浪费，虚弱和瘫痪。目前批准用于ALS的两种药物可以延长几个月的寿命，并且无法治愈患者。因此，显然需要新的药物来治疗ALS。ALS的特征在于神经元中由蛋白质和核糖核酸（RNA）组成的分子复合物（称为核糖核蛋白（RNP）复合物）的异常代谢。突变的、异常定位的或以其他方式失调的蛋白质获得异常的RNA结合特性并在结构上扰乱RNP复合物，导致ALS病理。同样，疾病相关的RNA可以建立未经许可的RNA-蛋白质相互作用，对细胞RNA代谢产生深远的影响。我们发现，一种称为NEAT1_2的特异性RNA诱导RNP复合物（称为paraspeckles）的形成，可能是ALS发病机制中的重要参与者。虽然在健康个体的神经元中不存在，但这种RNA在ALS运动神经元中积累。此外，许多调节NEAT1_2/paraspeckles的蛋白质受到ALS突变的影响。我们相信它有保护作用，帮助神经元存活。然而，目前没有NEAT 1_2靶向药物可用。传统的药物发现工作集中在蛋白质上，但最近已经有可能鉴定作用于RNA及其与蛋白质的复合物的药物。更具体地说，目前正在对患有致命性儿童疾病脊髓性肌萎缩症（SMA）的患者进行评估影响RNP的药物。最近，我们开发了一种系统，该系统将允许我们通过将NEAT1_2 RNA的一部分连接到多孔测试板中的微珠上来组装与NEAT1_2/paraspeckles组成相似的复合物。该系统可用于使用超快显微成像分析快速分析与这些复合物结合的蛋白质。在目前的提案中，我们的目标是更好地了解NEAT1_2/paraspeckles和ALS之间的联系，并找到可以调节这些复合物的新型小分子，这些复合物有可能用于ALS研究和药物发现。此外，我们将修改该系统，使其适合于发现小分子的任何ALS相关的RNA分子，使用一个主要的ALS致病RNA产生的C9 ORF 72基因。我们的团队在RNA生物学，ALS病理生物学和药物发现以及所有相关实验室工具方面具有重要的背景。因此，我们处于独特的位置，可以使用我们开发的上述最先进的方法，将我们对ALS病理生理学的理解转化为治疗开发并最终影响患者。