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.

核苷酸重复扩增突变会导致多种遗传性疾病，包括 1 型和 2 型强直性肌营养不良 (DM)，以及大部分额颞叶痴呆 (FTD) 和肌萎缩侧索硬化症 (ALS)。我们开发了一种基于细胞的筛查和二次验证分析。为了鉴定 Dm1 的小分子，我们筛选了多种小分子，并鉴定了 60 种新的命中化合物（强直性肌营养不良抑制剂 - MDI），这些化合物可用于二次测定。选择性地分散易于在 DM1 患者衍生细胞中形成核聚集体的扩增的含有 CUG 重复序列的 RNA，并隔离关键的 RNA 结合蛋白。值得注意的是，我们发现这些化合物还可以消散 DM2 患者衍生细胞中扩增的含有 CCUG 重复序列的 RNA 聚集点，这表明他们可能会进行治疗基于我们在 DM1 中获得的知识，研究了不同的富含 GC 的重复扩增疾病的潜力。在该项目中，我们设计了一种筛选策略，以确定由 C9orf72 基因 (C9FTD/ALS) 六核苷酸扩增引起的 FTD/ALS 的潜在小分子疗法。在该项目中，我们建议开发、优化和验证基于细胞的检测平台。接下来，我们将使用该平台来测试影响有毒 CUG RNA 焦点稳定性的化合物是否可以分散扩大的 G4C2 重复 RNA 焦点并减少二肽蛋白。最后，我们将对多种定制的小分子化合物进行概念验证筛选，以最好地评估我们筛选平台的价值和功效，以确定潜在的潜力。 C9FTD/ALS 的治疗剂。 FTD或ALS尚无有效治疗方法，鉴定具有潜在治疗价值的小分子为患有这些使人衰弱和致命的疾病的人带来希望。