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

对于由无义突变引起的疾病的治疗存在巨大需求。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一个有用的背景，因为CF是一个良好的 特征性遗传疾病，有许多模型系统，并有几十个无义突变内， CFTR。无义介导的衰变（NMD）是降解含有以下物质的转录物的细胞过程： 由无义突变引起的提前终止密码子。NMD在无稽之谈中充当治疗障碍 通过阻止蛋白质被翻译，因为细胞中几乎没有无义转录物，从而抑制突变条件。 然而，NMD并不平等地影响所有无意义的转录本。NMD的效率被认为会因 组织、细胞类型，甚至患者。此外，无义突变的特定位置内的 逃离国家导弹防御系统。该提案旨在研究CF无义突变中NMD的异质性， CF无义突变的药理学校正。含有CF无义的新型小鼠模型 预测逃脱NMD的突变将被开发出来。使用这些抗NMD模型，NMD效率将 测量并与含有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 研究了将研究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（其促进CFTR蛋白质折叠， 功能）将在小鼠和人类肠道类器官中单独和组合进行测试， 最终在体内无义突变小鼠模型中。最佳药理学组合可能因 每个基因型或NMD表型。研究NMD在CF异质性和CF无意义中的作用 突变校正可以阐明更好的治疗策略，并为追求精确的治疗提供证据。 医学方法在CF。本提案中概述的培训计划将有力地支持成功的过渡 转化研究的职业生涯。培训计划包括与科学有关的活动 通信、临床和转化研究、临床前体内动物技术和生物信息学。

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.