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

对无义突变引起的疾病的治疗是非常有必要的。在囊状组织的背景下 由CFTR基因突变引起的纤维化(CF)，具有两个无意义等位基因的患者缺乏CFTR特异性 治疗。Cf为研究无意义突变生物学提供了一个有用的环境，因为Cf是一口井 以遗传病为特征，有许多模型系统，并有数十个无意义的突变 CFTR。无意义介导的衰退(NMD)是细胞过程，降解转录本包含 无义突变导致的提前终止密码子。NMD在胡说八道中扮演着治疗障碍的角色 突变条件通过阻止蛋白质被翻译，因为细胞中只剩下很少的无意义转录本。 然而，NMD并不会平等地影响所有无稽之谈的记录。国家导弹防御系统的效率已经被建议通过 组织，细胞类型，甚至是病人。此外，基因中特定位置的无义突变 文字记录逃脱国家导弹防御系统。这项建议旨在研究Cf无义突变和NMD的异质性 无义突变的药理学矫正。含CF废话的新型小鼠模型 预测可以逃脱NMD的突变将被开发出来。使用这些抗NMD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。vbl.使用 现有和新的CF无义小鼠模型，药物纠正无义突变将是 学习。将研究NMD抑制剂以及读通剂(能够触发 提前终止密码子的通读)和CFTR调节子(它们促进CFTR蛋白折叠和 功能)将在小鼠和人类肠道器官中单独和联合测试 最终在体内进行无义突变的小鼠模型。最佳的药物组合可能不同于 每种基因型或由NMD表型决定。研究NMD在Cf异质性和Cf无稽之谈中的作用 突变校正可能阐明更好的治疗策略，并为追求精确度提供证据 CF中的医学方法。本提案中概述的培训计划将有力地支持成功过渡。 为实习生从事翻译研究。培训计划包括与科学有关的活动。 通信、临床和转译研究、临床前活体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.