Directed evolutioni of AAV for gene theraphy in a pig model of cystic fibrosis

用于囊性纤维化猪模型基因治疗的 AAV 定向进化

• 批准号: 7741474
• 负责人: Joseph Zabner
• 金额: $ 28.86万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7741474
• 关键词: Address; Aerosols; Affect; Alanine; Alveolus; Animal Model; Animals; Apical; Benchmarking; Binding; Birth; Capsid; Capsid Proteins; Cells; Chloride Ion; Chlorides; Clinical; Clinical Trials; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Dependovirus; Development; Disease; Disease Progression; Distal; Epithelial; Epithelium; Event; Evolution; Exhibits; Family suidae; Funding; Gene Delivery; Gene Transfer; Genes; Genetic; Genetic Recombination; Genome; Goals; Helper Viruses; Hereditary Disease; Human; Hybrids; Ileus; In Vitro; Infection; Instruction; Intestinal Obstruction; Intestines; Ion Transport; Knowledge; Lead; Leber' s amaurosis; Libraries; Life; Light; Lung; Lung diseases; Meconium; Mediating; Modeling; Morbidity - disease rate; Mutagenesis; Mutation; Natural Immunity; Neonatal; Newborn Animals; Newborn Infant; Nose; Organ; Pathogenesis; Pathway interactions; Patients; Phenotype; Process; Proteins; Pulmonary Cystic Fibrosis; Research; Respiratory Failure; Safety; Serotyping; Sialic Acids; Single-Stranded DNA; Specificity; Surface; Symptoms; Syndrome; Testing; Therapeutic; Threonine; Transgenes; Tropism; Viral; Virus; acquired immunity; airway epithelium; airway inflammation; airway surface liquid; clinical efficacy; clinical phenotype; cystic fibrosis airway epithelia; cystic fibrosis patients; disease-causing mutation; early cystic fibrosis; gene therapy; improved; in utero; in vitro Model; in vivo; macrophage; mortality; mutant; new therapeutic target; novel; prevent; programs; promoter; respiratory; tool; vector

Cystic fibrosis (CF) is a genetic disease caused by loss of cystic fibrosis transmembrane conductance regulator (CFTR). The disease involves many organs, and lung disease is the current major cause of morbidity and mortality. Gene transfer offers the potential to express CFTR in the lungs of patients and thereby slow or prevent disease progression. Yet despite outstanding research progress, we still lack answers to many crucial questions. A major impediment to progress has been the lack of an animal model that replicates disease typically found in humans. To circumvent limitations of current animal models, we have produced CFTR-/- and CFTRAF/- pigs. Newborn animals exhibit defective CI' transport and replicate abnormalities in newborn humans. Our preliminary data suggest that CFTR-/- pigs may also develop respiratory disease like humans. We will use this novel model to address questions key to CF. Aim 1. Will intestinal expression of CFTR prevent meconium ileus in CFTRAF508/- pigs? To answer this question, we will generate CFTRAF508/- pigs carrying a transgene expressing CFTR in the intestine. The resulting animals may be of value to other projects in the program, and the results may shed light on the underlying pathogenesis of meconium ileus and distal intestinal obstruction syndrome. Aim 2. Do genetic modifiers influence the clinical phenotype of CFTRAF508/AF508 pigs? We will cross our pigs to genetically diverse strains of pigs and ask how the CF phenotypes are altered. Discovering phenotypic changes could help identify pathways that modify the AF508 mutant protein or affect manifestations of the disease, thereby revealing novel therapeutic targets. Aim 3. When is CFTR expression required to alter the CF phenotype? We will generate CFTRAF508 pigs that express CFTR under control of an inducible promoter to answer key questions. Will CFTR expression in utero prevent disease in newborns? Will epithelial CFTR expression that begins after birth prevent lung disease? Will CFTR expression treat or slow the progression of established ainway disease? The answers will serve as a benchmark to guide therapeutic strategies including gene transfer. RELEVANCE (See instructions): Research in cystic fibrosis (CF) has been impeded by lack of an animal model that replicates the typical manifestations of the disease. Our Program recently developed a porcine model of CF. We will use this novel model to better understand CF and hopefully accelerate the discovery and testing of gene transfer and other novel preventions and treatments.

囊性纤维化（CF）是一种由囊性纤维化跨膜传导调节因子（CFTR）缺失引起的遗传性疾病。该疾病涉及许多器官，并且肺部疾病是目前发病率和死亡率的主要原因。基因转移提供了在患者肺部表达CFTR的可能性，从而减缓或预防疾病进展。然而，尽管研究取得了显著进展，我们仍然缺乏许多关键问题的答案。进展的一个主要障碍是缺乏一种复制人类典型疾病的动物模型。为了规避当前动物模型的局限性，我们已经产生了CFTR-/-和CFTRAF/-猪。新生动物在新生人类中表现出有缺陷的CI'转运和复制异常。我们的初步数据表明，CFTR-/-猪也可能像人类一样患上呼吸道疾病。我们将使用这个新的模型来解决CF的关键问题。 目标1。CFTR的肠道表达是否会预防CFTRAF 508/-猪的胎粪性肠梗阻？为了回答这个问题，我们将产生在肠道中携带表达CFTR的转基因的CFTRAF 508/-猪。由此产生的动物可能对该项目中的其他项目有价值，并且结果可能揭示胎粪性肠梗阻和远端肠梗阻综合征的潜在发病机制。目标二。遗传修饰因子是否影响CFTRAF 508/AF 508猪的临床表型？我们将把我们的猪与基因多样的猪种杂交，并研究CF表型是如何改变的。发现表型变化可以帮助确定修饰AF 508突变蛋白或影响疾病表现的途径，从而揭示新的治疗靶点。目标3.何时需要CFTR表达来改变CF表型？我们将产生在诱导型启动子控制下表达CFTR的CFTRAF 508猪，以回答关键问题。CFTR在子宫内的表达能预防新生儿疾病吗？出生后开始的上皮CFTR表达能预防肺部疾病吗？CFTR的表达会治疗或减缓已建立的尿路疾病的进展吗？这些答案将作为指导包括基因转移在内的治疗策略的基准。 相关性（参见说明）： 囊性纤维化（CF）的研究一直受到缺乏动物模型的阻碍， 疾病的典型表现。我们的项目最近开发了一个CF的猪模型。我们将使用这种新的模型来更好地了解CF，并希望加速基因转移和其他新的预防和治疗方法的发现和测试。