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

CFTR 对 CF 猪模型鼻窦和颅面发育的影响

• 批准号: 8527504
• 负责人: Eugene H Chang
• 金额: $ 12.78万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-02 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8527504
• 关键词: 3-Dimensional; Adult; Adverse effects; Affect; Animal Model; Antibiotics; Architecture; Bacteria; Birth; Caucasians; Caucasoid Race; Chloride Ion; Chlorides; Chronic; Chronic Sinusitis; Craniofacial Abnormalities; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Defect; Dental; Development; Disease; Electrophysiology (science); Environment; Environmental Risk Factor; Epithelial; Epithelium; Ethics; Exhibits; Face; Family suidae; Foundations; Future; Gallbladder; Genes; Goals; Growth; Health; Human; Image; Imaging technology; In Vitro; Infection; Inflammation; Intestines; Investigation; Knock-out; Lead; Liver; Lung; Lung diseases; Malnutrition; Maxillary Sinus; Modeling; Monitor; Morbidity - disease rate; Mutation; Organ; Pancreas; Paranasal Sinus Diseases; Pathogenesis; Pathology; Patients; Phenotype; Prevention; Regulator Genes; Research; Research Personnel; Respiratory physiology; Role; Sinus; Sinusitis; Site; Sphenoidal sinus; Sweat Glands; Techniques; Testing; Time; Work; airway epithelium; craniofacial; cystic fibrosis patients; disorder prevention; frontal sinus; gene therapy; improved; in utero; in vivo; interest; longitudinal human study; male; mortality; novel; prevent; reproductive

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.

描述（由申请人提供）：研究者的总体目标是开发一种动物模型来测试假设，以研究气道上皮细胞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。这是我们第一次通过动物模型研究囊性纤维化跨膜电导调节器 (CFTR) 对 CFTR 敲除猪 (CFTR-/-) 和野生型 (CFTR+/+) 猪的颅面发育和鼻窦病理的作用。囊性纤维化猪模型是第一个大型CF动物模型，我们的初步研究表明CF猪会出现鼻窦炎，并且颅面部变化与人类CF相似。我们将利用先进的成像技术和 3 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将发生变化。已知患有囊性纤维化的人类患者患有慢性鼻窦疾病以及鼻旁窦发育不全。 CF 患者通常有上颌窦发育不全，以及额窦和蝶窦较小或缺失。由于CF患者往往患有慢性鼻窦疾病，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，或者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时和整个成年期没有疾病的情况下研究鼻​​窦和颅面发育。具体目标 2：CFTR-/- 猪会先于肺部疾病发生鼻窦疾病。患有囊性纤维化的人类患者会出现鼻窦和肺部疾病。临床医生研究了鼻窦疾病对囊性纤维化患者肺部疾病进展的影响。对于鼻窦疾病和肺部疾病哪个先进行的研究尚未在人类中得到解答。如果鼻窦疾病确实先于肺部疾病发生，那么预防鼻窦疾病可以改善肺功能并降低 CF 的发病率和死亡率。 CF 猪将使我们能够在清洁的环境中以及细菌攻击后监测鼻窦和肺部疾病的发展。具体目标 3：CFTR-/- 猪将出现氯离子转运受损的上皮缺陷。我们可以通过对上皮细胞进行 CFTR 基因治疗来纠正这种缺陷。 CF 上皮细胞的体外气道培养物在电生理学研究中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 传递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猪将使我们能够针对鼻窦进行基因治疗。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。