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

基本信息

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
项目类别：
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

项目摘要

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.

对于废话突变引起的疾病非常需要治疗。在囊性的背景下由CFTR基因突变引起的纤维化（CF），两个废话等位基因的患者缺乏CFTR特异性治疗。 CF提供了一个有用的背景，在其中研究胡说八道的生物学，因为CF是一个井特征是遗传疾病，具有许多模型系统，并且在 CFTR。废话介导的衰减（NMD）是降解包含转录本的细胞过程过早的终止密码子是由胡说八道引起的。 NMD充当废话的治疗障碍突变条件是通过防止蛋白质被翻译而来的，因为几乎没有胡说八道的转录物保留在细胞中。但是，NMD并没有平等影响所有胡说八道的转录本。 NMD效率已被建议通过组织，细胞类型，甚至是患者。此外，在特定位置的废话突变成绩单逃脱NMD。该提案旨在研究CF胡说八道突变和 CF废话突变的药理校正。包含CF废话的新型鼠标模型预计将逃脱NMD的突变将开发。使用这些NMD抗性模型，NMD效率将可以测量并与现有的CF模型进行比较，该模型含有NMD易感废话突变。使用现有和新的CF胡说制小鼠模型，对胡说突变的药理学校正将是研究。 NMD抑制剂将进行研究，以及读取药物（能够触发过早停止密码子和CFTR调制器（促进CFTR蛋白折叠和功能）将独立测试，并在小鼠和人类肠癌中组合进行测试最终在胡说八道突变小鼠模型中体内。最好的药理组合可能有所不同每个基因型或NMD表型。研究NMD在CF异质性和CF废话中的作用突变校正可以阐明更好的治疗策略，并为追求精度提供证据 CF中的医学方法。该提案中概述的培训计划将强烈支持成功的过渡从事学员转化研究的职业。培训计划包括与科学有关的活动交流，临床和转化研究，体内动物技术和生物信息学。