Investigating the role of NMD in Nonsense Mutation Biology and Therapeutic Strategies

研究 NMD 在无义突变生物学和治疗策略中的作用

• 批准号: 10533441
• 负责人: Margaret Michicich
• 金额: $ 4.68万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533441
• 关键词: Alleles; Animal Model; Animals; Bioinformatics; Biological Assay; Biological Models; Biology; Caffeine; Cell physiology; Cells; Clinical Research; Clinical Trials; Communication Research; Congenic Mice; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Development; Disease; Drug Combinations; Duchenne muscular dystrophy; Embryo; Failure; Forskolin; Future; Genes; Genetic Diseases; Genotype; Heterogeneity; Heterozygote; Human; Human Genetics; In Vitro; Inbred CFTR Mice; Intestines; Lead; Location; Low Prevalence; Measures; Modeling; Mus; Mutation; Nonsense Codon; Nonsense Mutation; Nonsense-Mediated Decay; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phenotype; Process; Proteins; Resistance; Role; Science; Severities; Severity of illness; Small Intestines; Swelling; Techniques; Terminator Codon; Testing; Therapeutic; Tissues; Toxic effect; Training; Transcript; Translating; Translational Research; Translations; Treatment Efficacy; Work; amlexanox; antibiotic G 418; beta Thalassemia; burden of illness; career; cell type; cystic fibrosis mouse; cystic fibrosis patients; drug testing; efficacy testing; human tissue; improved; in vitro Model; in vivo; in vivo Model; in vivo evaluation; inhibitor; mouse model; mutation correction; novel; pre-clinical; precision medicine; premature; prevent; protein folding; protein function; resistance mutation; restoration; treatment response

There is a great need for treatments for diseases caused by nonsense mutations. In the context of Cystic Fibrosis (CF), caused by mutations in the CFTR gene, patients with two nonsense alleles lack CFTR-specific treatments. CF provides a useful context in which to study nonsense mutation biology as CF is a well characterized genetic disease, has numerous model systems, and has dozens of nonsense mutations within CFTR. Nonsense-mediated decay (NMD) is the cellular process that degrades transcripts containing premature termination codons caused by nonsense mutations. NMD acts as a treatment barrier in nonsense mutation conditions by preventing protein from being translated as few nonsense transcripts remain in the cell. NMD, however, does not affect all nonsense transcripts equally. NMD efficiency has been suggested to vary by tissue, cell type, and even by patient. Furthermore, nonsense mutations in specific locations within the transcript escape NMD. This proposal aims to study the heterogeneity of NMD in CF nonsense mutations and the pharmacological correction of CF nonsense mutations. Novel mouse models containing CF nonsense mutations predicted to escape NMD will be developed. Using these NMD resistant models, NMD efficiency will be measured and compared to existing CF models containing NMD susceptible nonsense mutations. Using existing and new CF nonsense mouse models, pharmacological correction of nonsense mutations will be studied. NMD inhibitors will be investigated, as well as readthrough agents (capable of triggering the readthrough of premature stop codons), and CFTR modulators (which promote CFTR protein folding and function) will be tested independently and in combination in mouse and human intestinal organoids and ultimately in vivo in nonsense mutation mouse models. The best pharmacological combination may differ for each genotype or by NMD phenotype. Studying the role of NMD in CF heterogeneity and CF nonsense mutation correction may elucidate better therapeutic strategies and provide evidence for pursuing a precision medicine approach in CF. The training plan outlined in this proposal will strongly support a successful transition to a career in translational research for the trainee. The training plan includes activities relating to science communication, clinical and translational research, preclinical in vivo animal techniques, and bioinformatics.

非常需要治疗由无义突变引起的疾病。在囊性的背景下 纤维化（CF），由 CFTR 基因突变引起，具有两个无义等位基因的患者缺乏 CFTR 特异性 治疗。 CF 为研究无义突变生物学提供了有用的背景，因为 CF 是一个很好的工具 特征性遗传疾病，有许多模型系统，并且内部有数十个无义突变 CFTR。无义介导的衰变（NMD）是降解包含以下内容的转录本的细胞过程： 由无义突变引起的过早终止密码子。 NMD 充当无意义的治疗障碍 由于细胞中几乎没有无义转录物，因此阻止蛋白质翻译，从而产生突变条件。 然而，NMD 并不同等地影响所有无意义转录本。建议 NMD 效率因人而异 组织、细胞类型，甚至患者。此外，在特定位置的无义突变 转录本逃逸 NMD。本提案旨在研究CF无义突变中NMD的异质性和 CF 无义突变的药理学纠正。含有 CF 废话的新型小鼠模型 预计将产生逃避 NMD 的突变。使用这些 NMD 抗性模型，NMD 效率将 进行测量并与包含 NMD 易感无义突变的现有 CF 模型进行比较。使用 现有的和新的 CF 无义小鼠模型，无义突变的药理学校正将是 研究过。将研究 NMD 抑制剂以及通读剂（能够触发 提前终止密码子的通读）和 CFTR 调节剂（促进 CFTR 蛋白折叠和 功能）将在小鼠和人类肠道类器官中进行独立和组合测试 最终在无义突变小鼠模型中进行体内实验。最佳药理学组合可能因不同情况而异 每个基因型或 NMD 表型。研究 NMD 在 CF 异质性和 CF 废话中的作用 突变校正可能阐明更好的治疗策略，并为追求精准治疗提供证据 CF 的医学方法。本提案中概述的培训计划将有力地支持成功过渡 为实习生从事转化研究的职业生涯。培训计划包括与科学相关的活动 交流、临床和转化研究、临床前体内动物技术和生物信息学。