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.

总结/摘要 囊性纤维化（CF）是一种遗传性疾病，影响美国约30，000人， 然后全世界有7万人CF是由上皮氯离子通道CF跨膜突变引起的 电导调节因子（CFTR）基因。在CF中，CFTR中的功能缺失突变减少了来自 细胞，并通过一种机制，提高上皮钠通道（ENaC）的活性，这是不完全的 明白这导致钠和水的重吸收增加，最终导致脱水。 上皮表面和在多个产生粘蛋白的器官，如肺， 鼻窦、肠、胰腺和生殖器官。CF患者在所有这些粘液中都出现临床症状- 生产器官。特别是，大多数CF患者的寿命缩短，因为CFTR的损失， 呼吸道，而且在疾病进展的早期发展肠道表型。这些肠道表型 与CF的肺表型相比，研究较少，但仍显著影响CF患者的生活。虽然CF是由 由于CFTR的许多不同突变，CFTR功能的差异不能解释患者的差异。 症状这表明CF的许多临床表型受遗传修饰剂和/或 环境因素这些遗传修饰剂和环境因素可能是开发的额外目标 治疗CF，可用于治疗所有患者，无论他们携带的突变。但不少 的潜在遗传修饰CF没有得到很好的研究和机制，他们修改CF 表型未知。 我们的实验室最近鉴定了一个果蝇CFTR基因的直系同源物，并建立了一个CF模型， 蝇肠上皮除了观察CFTR突变果蝇肠道上皮中的CF表型外， 发现了一种微小RNA，mir 263 a，作为ENaC活性的负调节因子。有趣的是， miR 263 a在CFTR突变果蝇中减少，表明CFTR对ENAC的调节受到调节， 部分地由MIR 263 A。在这里，我建议进一步描述果蝇突变模型的病理学特征，包括 研究细菌如何在该模型中调节疾病表型。然后我将使用飞CF模型获得 ENaC是一种已知的CF表型修饰剂。最后，由于寿命短，成本低， 果蝇的易处理性使其成为进行遗传筛选的理想模式生物，我建议识别新的 CF的潜在遗传修饰剂。总之，本工作将建立果蝇作为一个有用的模型，研究CF 并潜在地为疾病的治疗提供新的分子靶点。