Discovery of novel mechanisms that impact CFTR translation and contribute to cystic fibrosis pathogenesis

发现影响 CFTR 翻译并导致囊性纤维化发病机制的新机制

基本信息

批准号：
10545091
负责人：
JOHN L HARTMAN
金额：
$ 55.15万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-12-15 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10545091
关键词：
Acceleration Address Biogenesis Biological Models Birth Caucasians Cell model Cells Codon Nucleotides Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Defect Disease Engineering Eukaryota Genetic Diseases Genetic Models Genetic Polymorphism Goals Homologous Gene Human Individual Inherited Ion Transport Knowledge Learning Left Libraries Link Mammalian Cell Maps Mediating Mediator Medical Messenger RNA Methods Modeling Modification Molecular Molecular Abnormality Molecular Target Mutation Other Genetics Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Process Protein Biosynthesis Refractory Research Design Ribosomes Risk Role Saccharomyces cerevisiae System Technology Testing Transcript Translations Variant Work Yeasts airway epithelium autosome bioelectricity cellular targeting clinical phenotype clinically relevant clinically significant deep sequencing disease phenotype gene interaction genome-wide genome-wide analysis human disease improved in vivo induced pluripotent stem cell innovation insight molecular phenotype monolayer novel personalized medicine pharmacologic phenomics posttranscriptional premature protein folding protein misfolding ribosome profiling social tool translational impact

项目摘要

Discovery of novel mechanisms that impact CFTR translation and contribute to cystic fibrosis pathogenesis Abstract This project is intended to advance high throughput yeast phenomics technology and provide new mechanistic information regarding a model genetic disease, cystic fibrosis (CF). When clinically significant mutations in the CF transmembrane conductance regulator (CFTR) are engineered in a yeast homolog (YOR1), genome-wide phenomic analysis using the yeast deletion strain library (YDSL) identifies novel pathways that impact CF molecular phenotype. New experimental targets for CF therapy also result from studies of this type. Under Specific Aim 1 of the renewal, we show ways in which genome-wide yeast studies can advance understanding of CFTR variants including nonsense (Type 1) and severe folding (Type 2) defects for which no “personalized” modulator treatment currently exists. We also demonstrate phenomics can be applied to elucidate pathways necessary for pharmacologic rescue. Specific Aim 2 engages leading-edge technology (ribosomal profiling, deep-sequencing based on modification mapping (Psi-Seq)) to delve deeply into novel (and unexpected) mediators of CFTR biogenesis based on analysis of mammalian translational efficiency. In particular, we examine the role of ribosomal “collisions” or “queueing” described by phenomic analysis, and mRNA pseudouridinylation for effects on CFTR translation and protein synthesis. Specific Aim 3, applies innovative CF cell models to establish clinical significance of findings from Aims 1 and 2. The proposed studies will test important hypotheses relevant to CF pathogenesis, provide new knowledge regarding translational velocity and mRNA utilization during CFTR expression, and show that ribosomal collisions and pseudouridinylation contribute to cystic fibrosis. Our findings will also help advance new targets for treating refractory forms of CF. Lessons learned from the studies can be applied to many other inherited and serious human illnesses.

发现影响CFTR翻译并有助于囊性纤维化的新型机制发病抽象的该项目旨在提高高通量酵母菌现象技术并提供新的机械有关模型遗传疾病，囊性纤维化（CF）的信息。当临床上显着的突变 CF跨膜电导调节剂（CFTR）在酵母同源物（YOR1），全基因组中设计使用酵母缺失应变文库（YDSL）的现象分析确定了影响CF的新途径分子表型。 CF治疗的新实验靶标也来自对这种类型的研究。在下面续订的特定目的1，我们展示了全基因组酵母研究可以提高理解的方式 CFTR变体包括胡说八道（1型）和严重的折叠（类型2）缺陷，没有“个性化” 当前存在调节剂治疗。我们还证明现象学可以应用于阐明途径药学救援所必需的。特定的目标2参与领先技术（核糖体分析，基于修改映射（PSI-SEQ）的深入序列，深入研究新颖（和意外） CFTR生物发生的介体基于哺乳动物翻译效率的分析。特别是我们检查现象分析描述的核糖体“碰撞”或“排队”和mRNA的作用假素胺化对CFTR翻译和蛋白质合成的影响。特定目标3，适用创新 CF细胞模型从AIM 1和2建立发现的临床意义。拟议的研究将测试与CF发病机理有关的重要假设，提供有关翻译速度和的新知识 CFTR表达过程中的mRNA利用率，并表明核糖体碰撞和假氨基化有助于囊性纤维化。我们的发现还将有助于促进治疗CF难治形式的新目标。从研究中汲取的教训可以应用于许多其他遗传和严重的人类疾病。