Investigation of CFTR on sinus and craniofacial development in a CF porcine model

描述（由申请人提供）：研究者的总体目标是建立一个动物模型，以验证研究气道上皮在颅面发育中的作用的假设，并了解囊性纤维化（CF）中窦性疾病的病理。这是动物模型首次允许我们研究囊性纤维化跨膜传导调节剂（CFTR）在CFTR敲除（CFTR-/-）和野生型（CFTR+/+）猪颅面发育和鼻窦病理中的作用。囊性纤维化猪模型是CF的第一个大型动物模型，我们的初步研究表明CF猪发生鼻窦炎，颅面变化与人类CF相似。我们将利用先进的成像技术和三维体积分析来研究WT和CF猪鼻窦疾病和颅面生长的变化。我们还具有用细菌挑战WT和CF猪的优势，并观察感染是否会导致猪的鼻窦疾病和颅面变化。我们假设CFTR的缺乏会导致猪CF模型颅面发育和鼻窦病理的变化。为了研究这一假设，我们提出了三个具体目标。具体目标1:CFTR-/-猪将有鼻窦和颅面发育的变化。人类囊性纤维化患者已知有慢性鼻窦疾病以及副鼻窦发育不全。CF患者通常有上颌窦发育不全，额窦和蝶窦小或缺失。由于CF患者常伴有慢性鼻窦疾病，一直难以确定是CF的遗传缺陷导致鼻窦发育不全导致鼻窦疾病，还是鼻窦疾病导致鼻窦发育不全，或者两者是独立的因素。CF猪将使我们能够在出生时和整个成年期没有疾病的情况下研究鼻窦和颅面发育。特异性目标2:CFTR-/-猪在肺部疾病之前会出现鼻窦疾病。患有囊性纤维化的人类患者会出现鼻窦和肺部疾病。临床医生已经研究了窦性疾病在囊性纤维化患者肺部疾病进展中的作用。鼻窦疾病和肺部疾病孰先孰后的研究还没有在人类中找到答案。如果窦性疾病确实先于肺部疾病，那么预防窦性疾病可以改善肺功能，降低CF的发病率和死亡率。CF猪将使我们能够在清洁的环境和细菌攻击后监测窦性疾病和肺部疾病的发展。特异性目标3:CFTR-/-猪将有氯离子运输受损的上皮缺陷。我们可以通过CFTR对上皮细胞的基因治疗来纠正这一缺陷。体外培养的CF上皮在电生理学研究中表现出氯化物运输缺陷。我们假设我们可以通过基因治疗将CFTR输送到气道上皮细胞来纠正这一缺陷。如果这在体外成功，我们就可以过渡到体内猪模型。CF猪将允许我们对鼻窦进行靶向基因治疗。这项工作将为未来治疗和预防人类鼻窦疾病奠定基础，并可能改善CF患者的肺功能和健康状况。

DESCRIPTION (provided by applicant): The overall goal of the investigator is to develop an animal model to test hypotheses that will study the role of airway epithelia in craniofacial development and understand the pathology of sinus disease in Cystic fibrosis (CF). This is the first time an animal model will allow us to investigate the role of Cystic Fibrosis Transmembrane conductance Regulator (CFTR) on craniofacial development and sinus pathology in CFTR knockout (CFTR-/-) and wild-type (CFTR+/+) pigs. The cystic fibrosis pig model is the first large animal model for CF, and our preliminary studies show that the CF pig develops sinusitis, and craniofacial changes similar to human CF. We will utilize advances in imaging technology and 3-dimensional volumetric analysis to investigate changes in paranasal sinus disease and craniofacial growth in WT and CF pigs. We also have the advantage of challenging both WT and CF pigs with bacteria, and observing if infection causes sinus disease and craniofacial changes in pigs. We hypothesize that a lack of CFTR will lead to changes in craniofacial development and sinus pathology in a porcine CF model. To investigate this hypothesis we propose three specific aims. Specific Aim 1: CFTR-/- pigs will have changes in sinus and craniofacial development. Human patients with cystic fibrosis are known to have chronic sinus disease as well as paranasal sinus hypoplasia. Patient with CF typically have maxillary sinus hypoplasia, as well as small or absent frontal and sphenoid sinuses. Because CF patients often have chronic sinus disease, it has been difficult to determine if the genetic defect of CF causes sinus hypoplasia leading to sinus disease, or sinus disease leads to sinus hypoplasia, or if they are independent factors. The CF pig will allow us to study sinus and craniofacial development in the absence of disease at birth and throughout adulthood. Specific Aim 2: CFTR-/- pigs will develop sinus disease prior to lung disease. Human patients with cystic fibrosis develop both sinus and lung disease. Clinicians have investigated the role of sinus disease to the progression of lung disease in humans with cystic fibrosis. The investigation of which comes first, sinus or lung disease has not been answered in humans. If sinus disease does precede lung disease, prevention of sinus disease may improve lung function and reduce morbidity and mortality in CF. The CF pig will allow us to monitor the development of sinus and lung disease in a clean environment and after bacterial challenge. Specific Aim 3: CFTR-/- pigs will have an epithelial defect of impaired chloride transport. We can correct this defect by gene therapy of CFTR to the epithelia. In vitro airway cultures of CF epithelia exhibit defective chloride transport in electrophysiology studies. We hypothesize that we can correct this defect by delivering CFTR to airway epithelia via gene therapy. If this is successful in vitro, we can transition to an in vivo pig model. The CF pig will allow us to target gene therapy in the sinus. This work will serve as a foundation for future treatment and prevention of sinus disease in humans, and may improve lung function and the health of CF patients.