CFTR AND AIRWAY PATHOLOGY OF CYSTIC FIBROSIS

在此续订申请中提出的研究试图了解 CFTR在人呼吸道上皮细胞功能中的作用及其机制 CF的功能障碍导致了其呼吸道病理特征 疾病。五个相关的项目将研究CFTR测距的各个方面 从分子到临床。Lingappa博士(项目1)将研究 CFTR是如何插入细胞膜并在细胞内运输的 手机。Verkman博士(项目2)将研究CFTR在 胞内细胞器。特别是，使用新开发的 技术，他将检验其他人的结论，即CFTR参与其中 在胞吐作用和调节内吞体内的pH方面。使用膜片钳 技术，Wine博士(项目3)将详细描述阴离子 野生型和突变型cftr的通道特性。来自我们的最新工作 中心已经证明，介体诱导的氯的分泌通过培养的 Cf呼吸道粘膜下腺细胞低于正常的5%。因此， 项目4(Widdicombe&Finkbeiner)的目标是检验假设 CF中最初的呼吸道粘液积聚主要反映了 腺体粘液分泌物脱水或其他改变。最后，Dr Szoka(项目5)将测试一种非病毒方法用于基因治疗 参见将尝试通过两种方法在体内转染呼吸道上皮细胞 雾化吸入和静脉注射。后一条路线增加了这种可能性 纠正粘膜下腺细胞的缺陷。所有项目都将利用 表面和腺上皮细胞的高分化原代培养 由芬克贝纳博士指导的细胞培养核心提供的细胞。

The proposed research in this renewal application seeks to understand what roles CFTR plays in the function of human airway epithelium, and how its malfunction in CF leads to the airway pathology characteristic of this disease. Five interrelated projects will study aspects of CFTR ranging from the molecular to the clinical. Dr. Lingappa (Project 1) will study how CFTR is inserted into cell membranes and is trafficked within the cell. Dr. Verkman (Project 2) will study the function of CFTR in intracellular organelles. In particular, using newly developed techniques, he will test the conclusions of others that CFTR is involved in exocytosis and in regulating the pH of endosomes. Using the patch-clamp technique, Dr. Wine (Project 3) will characterize in detail the anion channel properties of wild-type and mutant CFTR. Recent work from our Center has demonstrated that mediator-induced Cl secretion by cultures of CF airway submucosal gland cells is less than 5% of normal. Therefore, the aim of Project 4 (Widdicombe & Finkbeiner) will be to test the hypothesis that the initial accumulation of airway mucus in CF reflects predominantly dehydration or other alterations in gland mucous secretions. Finally, Dr Szoka (Project 5) will test a non-viral approach towards gene therapy of CF. Attempts will be made to transfect airway epithelium in vivo by both aerosol and intravenous routes. The latter route increases the likelihood of correcting defects in submucosal gland cells. All projects will utilize the highly differentiated primary cultures of surface and gland epithelial cells provided by a cell culture CORE directed by Dr Finkbeiner.