Correction Core

矫正核心

基本信息

批准号：
8249134
负责人：
SHERIF E GABRIEL
金额：
$ 15.88万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8249134
关键词：
Adherent Culture Adult Affect Animal Model Biological Assay Biopsy Caucasians Caucasoid Race Chronic Code Confocal Microscopy Cyclic AMP Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Defect Disease Dose Epithelial Epithelium Family suidae Gastrointestinal tract structure Gene Transfer Hereditary Disease Human Hydration status Ileus In Vitro Infection Intestines Ion Transport Ions Label Lead Liquid substance Lung Measurement Measures Meconium Mediating Molecular Mucous body substance Mus Neonatal Nose Operative Surgical Procedures Optics Organ Population Preparation Property Proteins Research Research Personnel Single-Gene Defect Surface Techniques Testing Thick Time Tissues Virus absorption airway epithelium airway surface liquid blue dextran cystic fibrosis airway cystic fibrosis airway epithelia cystic fibrosis mouse cystic fibrosis patients design effective therapy effectiveness measure electrical potential epithelial Na+ channel gastrointestinal epithelium gene cloning gene therapy gene transfer vector in vivo neonatal human neonate novel rectal vector

项目摘要

Cystic Fibrosis (CF) has been at the forefront of gene transfer research for the last decade. This disease, resulting from a single gene defect, affects the epithelia of multiple organs of the body including the lung and gastrointestinal tract. The lack of effective long-term treatment for the pulmonary manifestations of CF, the accessibility of the lung via the airway lumen, and the fact that viruses known to infect the lung were being developed into non-replicating gene transfer vectors led investigators to believe that administration of gene transfer vectors to the lung could potentially result in an effective treatment of this disease. The assessment of the efficacy of gene transfer for CF requires a routine and reliable assay to verify expression of functional CFTR. This Correction Core will provide multiple techniques to detect the efficacy of CFTR gene transfer at correcting the ion, fluid and mucus transport defects in both the airway and gut. Cultured airway epithelia from the CF human and mouse will be used extensively in Ussing chamber studies to demonstrate correction of the CFTR mediated Cl- secretion and Na+ hyperabsorption. These preparations will also be used to determine if the altered rate of fluid transport has been corrected by CFTR gene transfer using confocal microscopy and measurement of blue dextran absorption. The pH and mucus biophysical properties will be measured on CF cultured airway epithelia to ascertain CFTR correction. Both human and murine nasal PD assays will be provided to assess CFTR gene transfer in vivo. Ion transport across CF human rectal biopsy studied in Ussing chambers will also be available as will freshly excised epithelial tissue from neonatal murine airways and gut to measure the effectiveness of in vivo vector dosing in neonatal CF mice. Finally, in anticipation of a new animal model for CF (a CF pig) we have begun to characterize the bioelectric properties of the porcine epithelium so that we will be poised for those possible studies.

在过去的十年中，囊性纤维化（CF）一直处于基因转移研究的最前沿。这种疾病，由单个基因缺陷引起的影响人体多个器官的上皮，包括肺和胃肠道。缺乏对CF肺表现的有效长期治疗肺通过气道管腔的可及性，以及已知感染肺的病毒的事实正在发展为非复制基因转移载体，导致研究者相信基因的给药将向量转移到肺部可能会导致对该疾病的有效治疗。评估基因转移对CF的功效需要常规和可靠的测定以验证功能的表达 CFTR。该校正核心将提供多种技术来检测CFTR基因转移的功效纠正气道和肠道中的离子，流体和粘液转运缺陷。培养的气道上皮从CF人和小鼠中将广泛用于室内研究中，以证明校正CFTR介导的CL-分泌和Na+高吸收。这些准备工作也将是用于确定使用CFTR基因转移校正流体转运率是否改变的速率共聚焦显微镜和蓝色葡聚糖吸收的测量。 pH和粘液生物物理将在CF培养的气道上皮上测量特性，以确定CFTR校正。人类和将提供鼠鼻PD分析以评估体内CFTR基因转移。离子跨CF 在Ussing Charbers中研究的人类直肠活检也将提供新鲜切除的上皮组织从新生儿鼠气和肠道，以测量新生儿CF中体内载体剂量的有效性老鼠。最后，预计CF的新动物模型（CF猪）我们已经开始表征猪上皮的生物电特性，以便我们将有望进行这些可能的研究。