Drosophila as a model to study modifiers of Cystic Fibrosis

果蝇作为研究囊性纤维化修饰因子的模型

• 批准号: 10550126
• 负责人: Elizabeth Lane
• 金额: $ 7.43万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-03 至 2025-03-02
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10550126
• 关键词: Affect; Bacteria; Biological Assay; Candidate Disease Gene; Cell Line; Cells; Chloride Channels; Chlorides; Clinical; Colon; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Dehydration; Disease; Disease Progression; Drosophila genus; Environmental Risk Factor; Epithelial Cells; Epithelium; European; Genes; Genetic; Genetic Diseases; Genetic Screening; Germ-Free; Goals; Human; Intestinal Motility; Intestines; Ion Channel; Longevity; Lung; Messenger RNA; MicroRNAs; Modeling; Mucins; Mucous body substance; Mutation; Newborn Infant; Organ; Orthologous Gene; Other Genetics; Outcome; Pancreas; Pathology; Patients; Persons; Phenotype; Proliferating; Regulation; Regulator Genes; Respiratory System; Signal Pathway; Sinus; Sodium; Surface; Symptoms; Therapeutic; Time; Toxic effect; United States; Up-Regulation; Water; Work; absorption; autosome; clinical phenotype; clinically relevant; cohort; cost; cystic fibrosis patients; differential expression; disease phenotype; drug action; dysbiosis; epithelial Na+ channel; fly; gastrointestinal epithelium; inhibitor; insight; knock-down; loss of function mutation; microbiome; model organism; molecular targeted therapies; mutant; new therapeutic target; overexpression; recessive genetic trait; reproductive organ; sensor; therapeutic target; transcription factor

Summary/abstract Cystic Fibrosis (CF) is genetic disorder that effects approximately 30,000 people in the United states and more then 70,000 worldwide. CF is caused by mutations in the epithelial chloride channel CF transmembrane conductance regulator (CFTR) gene. In CF, loss-of-function mutations in CFTR, reduces chloride efflux from cells, and elevates the activity of the epithelial sodium channel (ENaC) through a mechanism that is not fully understood. This results in an increase sodium and water reabsorption, which ultimately leads to dehydration of the epithelial surface and reduction in mucus transport in multiple mucin-producing organs, such as the lungs, sinuses, intestine, pancreas, and reproductive organs. CF patients develop clinical symptoms in all these mucus- producing organs. In particular, most CF patients have shortened lifespans because of loss of CFTR in the respiratory tract, but also develop gut phenotypes early in the progression of the disease. These gut phenotypes are less studied than the lung phenotypes of CF but still significantly impact CF patients lives. While CF is caused by many different mutations in CFTR, the differences in CFTR function cannot explain the differences in patient symptoms. This indicates that many of the clinical phenotypes of CF are influenced by genetic modifiers and/or environmental factors. These genetic modifiers and environmental factors could be additional targets to develop treatments for CF that could be used to treat all patients regardless of the mutation they harbor. However, many of the potential genetic modifiers of CF are not well studied and the mechanism by which they modify CF phenotypes is unknown. Our lab has recently identified a Drosophila ortholog of the CFTR gene and established a CF model in the fly gut epithelium. In addition to observing CF phenotypes in the gut epithelium of CFTR mutant flies, we uncovered a micro RNA, mir263a, as a negative regulator of ENaC activity. Interestingly, the expression of mir263a is decreased in CFTR mutant flies, suggesting that that the regulation of ENAC by CFTR is regulated in part by mir263a. Here, I propose to further characterize the pathology of the fly mutant model including examining how bacteria can modulate disease phenotypes in this model. I will then use the fly CF model to gain new insight into ENaC, a known modifier of the CF phenotype. Finally, as the short lifespan, low cost, and genetic tractability of the fly makes it an ideal model organism to perform genetic screens, I propose to identify new potential genetic modifiers of CF. Altogether this work will establish Drosophila as a useful model to study CF and potentially provide new molecular targets for treatment of the disease.

摘要/摘要 囊性纤维化（CF）是遗传疾病，在美国影响约30,000人，更多 然后在全球70,000。 CF是由上皮氯化物通道CF跨膜突变引起的 电导调节剂（CFTR）基因。在CF中，CFTR的功能丧失突变，减少了氯化物外排 细胞，并通过不完全的机制提高上皮钠通道（ENAC）的活性 理解。这导致钠和水的吸收增加，最终导致 多个产生粘蛋白的器官（例如肺， 鼻窦，肠，胰腺和生殖器官。 CF患者在所有这些粘液中出现临床症状 - 产生器官。特别是，由于CFTR失去了CFTR，大多数CF患者的寿命缩短 呼吸道，但在疾病进展的早期也会发展出肠道表型。这些肠道表型 与CF的肺表型相比，研究较少，但仍然显着影响CF患者的生活。当CF引起 通过CFTR中许多不同的突变，CFTR功能的差异无法解释患者的差异 症状。这表明CF的许多临床表型都受遗传修饰剂和/或的影响 环境因素。这些遗传修饰符和环境因素可能是发展的其他目标 CF的治疗方法可用于治疗所有患者，而不管他们携带的突变如何。但是，很多 在CF的潜在遗传修饰符中，没有很好地研究其修改CF的机制 表型未知。 我们的实验室最近确定了CFTR基因的果蝇直系同源物，并在 飞肠上皮。除了观察CFTR突变体肠道上皮中的CF表型外，我们还 发现了微RNA，miR263a，作为ENAC活性的负调节剂。有趣的是，表达 CFTR突变蝇中的miR263a降低，表明调节CFTR的ENAC调节 部分由mir263a。在这里，我建议进一步表征蝇突变模型的病理 检查细菌如何在该模型中调节疾病表型。然后，我将使用Fly CF模型获得 对ENAC的新见解，ENAC是CF表型的已知修饰符。最后，作为寿命短，低成本和遗传 苍蝇的障碍性使其成为执行遗传筛查的理想模型生物，我建议识别新的 CF的潜在遗传修饰符。总共这项工作将建立果蝇作为研究CF的有用模型 并有可能为治疗该疾病的新分子靶标提供。