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

描述（由申请方提供）：研究者的总体目标是开发一种动物模型，以检验将研究气道上皮在颅面发育中的作用并了解囊性纤维化（CF）中鼻窦疾病的病理学的假设。这是第一次动物模型将使我们能够研究囊性纤维化跨膜传导调节因子（CFTR）对CFTR敲除（CFTR-/-）和野生型（CFTR+/+）猪的颅面发育和鼻窦病理学的作用。囊性纤维化猪模型是第一个大型的CF动物模型，我们的初步研究表明，CF猪发生鼻窦炎，颅面改变类似于人类CF。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猪的副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。我们假设缺乏CFTR将导致猪CF模型中颅面发育和鼻窦病理学的变化。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-/-猪的鼻窦和颅面发育会发生变化。患有囊性纤维化的人类患者已知患有慢性鼻窦疾病以及鼻窦发育不全。CF患者通常有上颌窦发育不全，以及小或缺乏额窦和蝶窦。由于CF患者通常患有慢性鼻窦疾病，因此很难确定CF的遗传缺陷是否导致鼻窦发育不全导致鼻窦疾病，或鼻窦疾病导致鼻窦发育不全，或者它们是否是独立因素。CF猪将使我们能够在出生和整个成年期无疾病的情况下研究鼻窦和颅面发育。具体目标2：CFTR-/-猪将在肺部疾病之前发生鼻窦疾病。患有囊性纤维化的人类患者会同时发生鼻窦和肺部疾病。临床医生研究了鼻窦疾病对囊性纤维化患者肺部疾病进展的作用。鼻窦或肺部疾病的研究在人类中尚没有答案。如果鼻窦疾病确实先于肺部疾病，那么预防鼻窦疾病可以改善CF的肺功能，降低发病率和死亡率。CF猪将使我们能够在清洁的环境中和细菌攻击后监测鼻窦和肺部疾病的发展。具体目标3：CFTR-/-猪将具有受损的氯离子转运的上皮缺陷。通过对上皮细胞进行CFTR的基因治疗可以纠正这一缺陷。在电生理学研究中，CF上皮细胞的体外气道培养物表现出氯离子转运缺陷。我们假设我们可以通过基因治疗将CFTR递送到气道上皮来纠正这种缺陷。如果这在体外是成功的，我们可以过渡到体内猪模型。CF猪将允许我们在鼻窦中进行靶向基因治疗。这项工作将作为未来治疗和预防人类鼻窦疾病的基础，并可能改善CF患者的肺功能和健康。 公共卫生相关性：鼻窦疾病和囊性纤维化（CF）颅面发育的发病机制尚不清楚。CF猪动物模型将允许我们研究CFTR在颅面和窦上皮中的作用。这些研究将作为治疗和预防鼻窦疾病的治疗方法的基础，并了解气道上皮细胞在颅面发育中的作用。

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. PUBLIC HEALTH RELEVANCE: The pathogenesis of sinus disease and craniofacial development in cystic fibrosis (CF) is unknown. The CF pig animal model will allow us to investigate the role of CFTR in craniofacial and sinus epithelia. These studies will serve as a foundation for therapies to treat and prevent sinus disease and understand the role of airway epithelia in craniofacial development.