REGULATION OF AIRWAY SUBMUCOSAL GLAND DEVELOPMENT

气道粘膜下腺发育的调节

• 批准号: 3083310
• 负责人: MICHAEL D INFELD
• 金额: $ 8.34万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3083310
• 关键词: RNase protection assay; antiserum; cell cell interaction; cell growth regulation; complementary DNA; extracellular matrix; fibroblasts; fibronectins; gene expression; histogenesis; human genetic material tag; immunocytochemistry; in situ hybridization; laboratory rabbit; laboratory rat; messenger RNA; phenotype; protein isoforms; respiratory epithelium; tissue /cell culture; transfection; western blottings

This project examines the role of fibronectin (Fn) in regulating epithelial invasion of the extracellular matrix (ECM) during airway submucosal gland (SMG) morphogenesis. Interest in specific, alternatively-spliced isoforms of the glycoprotein fibronectin as mediators of epithelial morphogenesis is supported by the developmental biology literature and preliminary data from our laboratory using cultured human cells which morphologically model early SMG development. Using immuno-histochemistry and in situ hybridization, active Fn isoforms will be localized in our cell culture model and neonatal rat SMGs in vivo. Transcriptional and translational regulation of specific Fn isoform expression in various human cell lines will be investigated by nuclease protection assay and immunoblotting. This will be correlated with the cell lines' ability to promote epithelial ECM invasion in vitro. Finally, cell lines constitutively unable to support epithelial invasion will be transfected with cDNA constructs from Fn domains specific for the isoforms identified as important to epithelial invasion. Transfected fibroblasts will be compared to untransfected controls to determine if expression of these sequences confers the ability to promote epithelial invasive behavior in vitro. These studies may have relevance beyond airway development for clinical problems such as repair after injury and tumor progression.

该项目检查了纤连蛋白（FN）在调节中的作用 气道期间细胞外基质（ECM）的上皮入侵 粘膜下腺（SMG）形态发生。 对特定的兴趣 或者，糖蛋白纤连蛋白的同工型或 上皮形态发生的介体得到了发展的支持 生物学文献和我们实验室的初步数据使用 培养的人类细胞形态上对早期SMG发育进行了建模。 使用免疫 - 归化学和原位杂交，主动FN同工型 将位于我们的细胞培养模型和新生大鼠SMG中 体内。 特定FN的转录和翻译调节 各种人细胞系中的同工型表达将通过 核酸酶保护测定和免疫印迹。这将是相关的 具有细胞系在体外促进上皮ECM侵袭的能力。 最后，细胞系数无法支撑上皮入侵 将用特有的FN域的cDNA构建体转染 同工型对上皮侵袭很重要。 转染 将成纤维细胞与未转染的对照进行比较，以确定是否是否 这些序列的表达赋予了促进上皮的能力 体外侵入性行为。 这些研究可能超出 临床问题的气道开发，例如受伤后修复和 肿瘤进展。