Development and validation of a cell-based assay system to identify novel therapeutics for C9ALS/FTD

开发和验证基于细胞的检测系统，以确定 C9ALS/FTD 的新疗法

基本信息

批准号：
9978295
负责人：
LUCIO COMAI
金额：
$ 45.38万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2023-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9978295
关键词：
ALS patients Affect Amyotrophic Lateral Sclerosis Animal Model Applications Grants Biological Assay Biotechnology C9ORF72 CUG repeat Cells Chemicals Clinical Collaborations Collection Custom Development Dipeptides Disease Frontotemporal Dementia Funding Future Genes Genetic Diseases Goals Hand Human Resources Individual Introns Knowledge Lead Libraries Measures Molecular Motor Neurons Mutation Myotonic Dystrophy Myotonic dystrophy type 1 Neurologist Nuclear Nucleotides Other Genetics Outcome Output Pathogenesis Pathologic Patients Pharmaceutical Preparations Pharmacology Property Protein Biosynthesis Proteins RNA RNA-Binding Proteins Research Specificity System Testing Therapeutic Therapeutic Agents Toxic effect Translations United States National Institutes of Health Validation Work base c9FTD/ALS candidate identification design drug candidate drug development effective therapy efficacy testing frontotemporal lobar dementia-amyotrophic lateral sclerosis in vivo inhibitor/antagonist interest nervous system disorder novel novel therapeutics programs screening small molecule small molecule libraries small molecule therapeutics therapeutic development

项目摘要

Nucleotide repeat expansion mutations cause a variety of genetic disorders including myotonic dystrophy (DM) types 1 and 2, and a large fraction of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). We developed a cell-based screen and secondary validation assays to identify small molecules for Dm1. We screened a diverse collection of small molecules and identified 60 novel hit compounds (myotonic dystrophy inhibitors-MDIs) that in secondary assays selectively disperse expanded CUG repeat- containing RNAs prone to form nuclear aggregates and sequester critical RNA-binding proteins in DM1 patient’s derived cells. Significantly, we showed that these compounds also dissipate expanded CCUG repeat-containing RNA foci in DM2 patient’s derived cells, suggesting that they may hold therapeutic potential across distinct GC-rich repeat expansion diseases. Building on the knowledge gained in our DM1 project, we have designed a screening strategy to identify potential small molecule therapeutics for FTD/ALS caused by an hexanucleotide expansion in the C9orf72 gene (C9FTD/ALS). In this project, wepropose to develop, optimize and validate a cell-based assay platform to measure critical molecular parameters of C9FTD/ALS disease. Next, we will use this platform to test whether compounds that affect the stability of toxic CUG RNA foci can disperse expanded G4C2 repeat RNA foci and decrease dipeptide proteins accumulation, two causes of toxicity in C9FTD/ALS patient’s derived cells. Lastly, we will perform a proof-of-concept screen of a diverse custom set of small-molecule compounds to best assess the value and efficacy of our screening platform to identify potential therapeutic agents for C9FTD/ALS. As there is no effective treatment for FTD or ALS, the identification of small molecules of potential therapeutic value offers hope for individuals with these debilitating and fatal diseases.

核苷酸重复膨胀突变引起多种遗传疾病，包括1和2型肌动物营养不良（DM），以及大部分额颞痴呆（FTD）和肌萎缩性侧向硬化症（ALS）。我们开发了一个基于细胞的筛选和二级验证测定法，以鉴定DM1的小分子。我们筛选了一个小分子的潜水员集合，并鉴定了60种新型HIT化合物（肌发育症抑制剂-MDIS），在次级测定中选择性地分散了膨胀的CUG重复重复 - 含有含有核聚集物的RNA，可形成核聚集物，并将临界RNA结合蛋白质在DM1患者中的细胞中。值得注意的是，我们表明这些化合物还耗散了DM2患者衍生细胞中的扩展的CCUG重复含RNA焦点，这表明它们可以持有治疗跨不同GC富含GC的重复扩张疾病的潜力。基于我们DM1中获得的知识项目，我们设计了一种筛选策略，以鉴定由C9ORF72基因（C9FTD/ALS）中的六核苷酸扩张引起的FTD/ALS的潜在小分子治疗。在这个项目中，Wepropose开发，优化和验证基于细胞的测定平台，以测量C9FTD/ALS疾病的关键分子参数。接下来，我们将使用该平台测试是否影响有毒CUG RNA焦点稳定性的化合物可以分散扩展的G4C2重复RNA焦点并减少二肽蛋白的积累，这是C9FTD/ALS患者衍生细胞中毒性的两个原因。最后，我们将对潜水员自定义的一组小分子化合物进行概念验证屏幕，以最好地评估筛选平台的价值和效率，以识别C9FTD/ALS的潜在治疗剂。如有没有有效的FTD或ALS处理，可以鉴定潜在治疗值的小分子为患有这些衰弱和致命疾病的人提供希望。