CFTR AND AIRWAY PATHOLOGY OF CYSTIC FIBROSIS

本更新申请中的拟议研究旨在了解 CFTR在人气道上皮功能中的作用，以及其如何 CF中的功能障碍导致这种气道病理学特征， 疾病五个相互关联的项目将研究CFTR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 cell. Verkman博士（项目2）将研究CFTR的功能， 胞内细胞器 特别是，利用新开发的 技术，他将测试其他人的结论，CFTR参与 在胞吐作用和调节内体的pH方面。使用膜片钳 技术，葡萄酒博士（项目3）将详细描述阴离子 野生型和突变型CFTR的通道特性。我们最近的工作 中心已经证明，介体诱导的Cl分泌的培养物， CF气道粘膜下腺细胞少于正常的5%。因此 项目4（Widdicombe & Finkbeiner）的目的是检验假设 CF中气道粘液的初始积聚主要反映了 脱水或腺体粘液分泌物的其他改变。最后，博士 Szoka（项目5）将测试一种非病毒的基因治疗方法， 参见 将尝试通过两种方法在体内切除气道上皮， 气雾剂和静脉途径。 后一种途径增加了 修复粘膜下腺细胞的缺陷。所有项目将利用 表面和腺上皮的高度分化的原代培养物 细胞由Finkbeiner博士指导的细胞培养核心提供。

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.