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

基本信息

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
项目类别：
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&apos 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

项目摘要

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）是由囊性纤维化跨膜电导调节剂（CFTR）丧失引起的遗传疾病。该疾病涉及许多器官，肺病是当前发病率和死亡率的主要原因。基因转移提供了表达患者肺中CFTR的潜力，从而缓慢或防止疾病进展。尽管取得了出色的研究进展，但我们仍然缺乏许多关键问题的答案。进步的主要障碍是缺乏复制人类通常发现的疾病的动物模型。为了规避当前动物模型的局限性，我们产生了CFTR - / - 和CFTRAF/ - 猪。新生动物表现出有缺陷的CI的运输和复制异常。我们的初步数据表明，CFTR - / - 猪也可能患有像人类一样的呼吸道疾病。我们将使用这种新颖的模型来解决CF的问题。 AIM 1。CFTR的肠道表达会预防CFTRAF508/ - 猪中的幼虫吗？为了回答这个问题，我们将生成CFTRAF508/ - 携带肠中带有转基因CFTR的猪。所产生的动物可能对程序中的其他项目具有价值，结果可能会阐明幼稚病的潜在发病机理和远端肠梗阻综合征。目标2。遗传修饰符是否会影响CFTRAF508/AF508猪的临床表型？我们将把猪跨到遗传上多样化的猪菌株，并询问如何改变CF表型。发现表型变化可以帮助识别改变AF508突变蛋白或影响疾病表现的途径，从而揭示新的治疗靶标。 AIM 3。改变CF表型需要CFTR表达何时？我们将生成CFTRAF508猪，该猪在控制诱导启动子的控制下表达CFTR，以回答关键问题。 CFTR在子宫内表达会预防新生儿疾病吗？出生后开始的上皮CFTR表达会预防肺部疾病吗？ CFTR表达会治疗或减慢已建立的AINWAY疾病的进展吗？答案将是指导包括基因转移在内的治疗策略的基准。相关性（请参阅说明）：缺乏复制动物模型，阻碍了囊性纤维化的研究（CF）该疾病的典型表现。我们的计划最近开发了CF的猪模型。我们将使用这种新颖的模型更好地理解CF，并希望加速基因转移以及其他新型预防和治疗的发现和测试。