MECHANISMS OF PULMONARY EPITHELIAL CELL DIFFERENTIATION

肺上皮细胞分化机制

• 批准号: 2519431
• 负责人: BRIAN P HACKETT
• 金额: $ 10.92万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519431
• 关键词: cell biology; cell cell interaction; cell differentiation; cell growth regulation; clone cells; early embryonic stage; flow cytometry; gel electrophoresis; genetic library; genetic markers; genetically modified animals; in situ hybridization; laboratory mouse; lung; mesenchyme; molecular biology; nucleic acid probes; polymerase chain reaction; respiratory epithelium; secretory protein; southern blotting; subtraction hybridization

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.

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，要么作为回应 肺损伤，肿瘤转化或固有异常 生长和分化是在 人肺病。 此外，对生物学的理解 肺上皮的生长和分化对于 意识到肺中基因疗法的全部潜力。 长期 这些研究的目的是定义分子和细胞 调节肺部近端生长和分化的机制 上皮。研究生长和分化机制 发展肺部将导致对这些作用的重要见解 过程在肺部疾病中发挥作用。 该提案的具体目的是 旨在检查开发中细胞的谱系关系 气道上皮和调节分化的分子机制 在这个上皮中。这些研究将利用独特的转基因 原始近端和远端肺上皮的标志物检查 早期肺部分配给不同谱系的细胞分配 发展。肺间隙在调节这些方面的作用 早期发育决定将在外植的胎儿肺中检查 正常间质上皮相互作用的破坏。 额外的 洞悉发育中细胞的谱系关系 肺上皮和克拉拉细胞作为祖细胞的作用将 通过表达克拉拉细胞分泌的细胞的毒素消融获得 蛋白-HSV胸苷激酶转基因在发育过程中。 最后， 近端肺部分化的潜在介体 上皮将被鉴定并以 减法杂交。荧光激活的细胞分选将是 用于获得原始近端和原始远端种群 表达β-半乳糖苷酶转基因的肺上皮细胞。 近端和远端上皮cDNA之后 库，将进行减法杂交以隔离和 表征来自原始肺部的独特克隆 上皮。