MECHANISMS OF PULMONARY EPITHELIAL CELL DIFFERENTIATION

肺上皮的生长和分化是必不可少的。 肺功能的决定因素。这些相同的过程，要么作为一种回应 肺损伤、肿瘤性转化或肺固有的异常 生长和分化是其发病机制中的重要因素 人类肺病。此外，对生物的了解 肺上皮细胞的生长和分化是 充分发挥肺部基因治疗的潜力。从长远来看 这些研究的目的是定义分子和细胞 近端肺组织生长和分化的调控机制 上皮组织。中华绒毛虫生长分化机制的研究 肺的发育将导致对这些角色的重要洞察 过程在肺部疾病中发挥作用。这项建议的具体目标是 旨在研究发育过程中细胞的谱系关系。 呼吸道上皮细胞及其调控分化的分子机制 在这层上皮内。这些研究将利用独特的转基因技术 原始近端和远端肺上皮标志物的检测 肺早期细胞向不同谱系的分配 发展。肺间充质细胞在调节这些功能中的作用 早期的发育决定将在移植的胎儿肺中进行检查 破坏正常间充质上皮的相互作用。其他内容 对发育过程中细胞的谱系关系的洞察 肺上皮细胞和Clara细胞作为祖细胞的作用 通过毒性消融表达Clara细胞分泌的细胞而获得 蛋白质-单纯疱疹病毒胸苷激酶基因在发育过程中的转基因。最后， 近端肺组织中潜在的分化介质 上皮细胞将通过技术鉴定和表征 消减杂交。荧光激活的细胞分选将被 用于获得原始近端和原始远端的种群 表达β-半乳糖苷酶转基因的肺上皮细胞。 构建近端和远端上皮细胞的c DNA 文库，将进行消减杂交以分离和 鉴定原始近端肺的独特克隆 上皮组织。

Growth and differentiation of the pulmonary epithelium are essential determinants of lung function. These same processes, either as a response to lung injury, neoplastic transformation, or intrinsic abnormalities of growth and differentiation, are important factors in the pathogenesis of human lung disease. Additionally, an understanding of the biology of growth and differentiation of the pulmonary epithelium is essential for realizing the full potential of gene therapy in the lung. The long term objective of these studies is to define the molecular and cellular mechanisms regulating growth and differentiation in the proximal pulmonary epithelium. A study of the mechanisms of growth and differentiation in the developing lung will lead to important insights into the role these processes play in lung disease. The specific aims in this proposal are intended to examine the lineage relationships of cells in the developing airway epithelium and the molecular mechanisms regulating differentiation within this epithelium. These studies will utilize unique transgenic markers of primitive proximal and distal pulmonary epithelium to examine the allocation of cells to different lineages during early lung development. The role of the pulmonary mesenchyme in regulating these early developmental decisions will be examined in explanted fetal lungs by disruption of normal mesenchymal epithelial interactions. Additional insights into the lineage relationships of cells in the developing pulmonary epithelium and the role of Clara cells as progenitor cells will be gained by toxigenic ablation of cells expressing a Clara cell secretory protein-HSV thymidine kinase transgene during development. Finally, potential mediators of differentiation in the proximal pulmonary epithelium will be identified and characterized by the technique of subtractive hybridization. Fluorescence activated cell sorting will be used to obtain populations of primitive proximal and primitive distal pulmonary epithelial cells expressing a beta-galactosidase transgene. Following the construction of proximal and distal epithelial cDNA libraries, subtractive hybridization will be performed to isolate and characterize unique clones from the primitive proximal pulmonary epithelium.