Targeted Inhibition of NMD to Enhance the Efficacy of Readthrough Drugs

靶向抑制NMD以增强通读药物的疗效

基本信息

批准号：
8536425
负责人：
Adrian R Krainer
金额：
$ 27.36万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8536425
关键词：
Accounting Age of Onset Age-Years Alleles Aminoglycoside Antibiotics Antisense Oligonucleotides Binding Bypass Cell Culture Techniques Cell Line Cells Complex Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Deposition Digestion Disease Drug usage Duchenne muscular dystrophy Effectiveness Exons Future Gene Expression Gene Proteins Genes Genetic screening method Goals Hereditary Disease Human Genome Project Immunoprecipitation In Vitro Individual Laboratories Lead Length Maps Mediating Messenger RNA Methods Micrococcal Nuclease Molecular Mutation Neonatal Nonsense Codon Nonsense Mutation Nucleotides Outcome Output Pathway interactions Patients Performance Pharmaceutical Preparations Production Proteins Quality Control RNA RNA Binding RNA Splicing Radioactive Reporter Rett Syndrome Reverse Transcriptase Polymerase Chain Reaction Ribosomes SMN2 gene Signal Transduction Spinal Muscular Atrophy Symptoms Technology Testing Therapeutic Transcript Translations base beta Thalassemia design effective therapy improved in vivo knock-down mRNA Decay mRNA Precursor mutant novel novel strategies phosphorothioate prevent research study tool

项目摘要

DESCRIPTION (provided by applicant): The successful completion of the human genome project and mapping of many disease genes, combined with advances in the molecular understanding of gene-expression pathways, provides unprecedented opportunities to design novel mechanism-based therapies for various genetic diseases. Nonsense mutations account for a large fraction of the causal mutations in nearly all genetic diseases. By definition, nonsense mutations introduce premature termination codons (PTCs), resulting in truncated proteins, and usually severe disease presentations. Translational readthrough drugs, such as ataluren, allow the synthesis of some full-length, functional protein from defective genes with nonsense mutations. However, nonsense-mediated mRNA decay (NMD)¿a ubiquitous mRNA quality-control pathway¿diminishes the effectiveness of readthrough drugs. We will selectively abrogate NMD of mRNAs harboring PTCs, so as to increase their availability for readthrough drugs. To explore the feasibility of our approach, we will initially focus on cell-culture experiments with several nonsense alleles of CFTR, MECP2, DMD, and HBB genes, which cause cystic fibrosis, Rett syndrome, Duchenne muscular dystrophy, and beta-thalassemia, respectively. We will systematically test the effect of inhibiting NMD on mRNA accumulation. Using reporter cell lines, we will then combine transcript-specific NMD inhibition with ataluren treatment, and determine whether there is increased synthesis of full-length protein, compared to ataluren treatment alone. The results of this exploratory study are expected to provide proof of principle for the effectiveness of targeted inhibition of NMD to enhance the efficacy of readthrough drugs. The proposed experiments could lead to a broadly applicable therapeutic approach that would be used in combination with readthrough drugs to treat a large number of severe genetic diseases.

描述（由申请人提供）：人类基因组计划的成功完成和许多疾病基因的定位，加上对基因表达途径的分子理解的进步，为设计针对各种遗传疾病的新型基于机制的疗法提供了前所未有的机会。无义突变占几乎所有遗传疾病的因果突变的很大一部分。根据定义，无义突变会引入过早终止密码子（PTC），导致蛋白质截短，并且通常会导致严重的疾病表现，例如转化药物。 ataluren 允许从具有无义突变的缺陷基因合成一些全长功能蛋白，然而，无义介导的 mRNA 衰减 (NMD)¿普遍存在的 mRNA 质量控制途径¿我们将选择性地消除含有 PTC 的 mRNA 的 NMD，以提高其对通读药物的可用性。为了探索我们方法的可行性，我们将首先关注 CFTR 的几个无义等位基因的细胞培养实验。、MECP2、DMD和HBB基因，分别导致囊性纤维化、雷特综合征、杜氏肌营养不良症和β地中海贫血，我们将系统地测试抑制效果。 NMD 对 mRNA 积累的影响。然后，我们将转录特异性 NMD 抑制与 ataluren 治疗相结合，并确定与单独的 ataluren 治疗相比，全长蛋白质的合成是否增加。这项探索性研究的结果是预期的。为靶向抑制 NMD 的有效性提供原理证明，以增强通读药物的功效。拟议的实验可能会产生一种广泛适用的治疗方法，该方法将与通读药物联合使用来治疗大量严重的遗传疾病。。