MUCIN EXPRESSION IN CYSTIC FIBROSIS & NORMAL EPITHELIUM

囊性纤维化中的粘蛋白表达

• 批准号: 2226950
• 负责人: Judith A Voynow
• 金额: $ 10.01万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-30 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2226950
• 关键词: bronchial mucus; chloride channels; clone cells; cystic fibrosis; gene expression; genetic transcription; genetic translation; glycoprotein biosynthesis; human subject; messenger RNA; mucins; respiratory epithelium; secretion; transfection

DESCRIPTION: (Adapted from the applicant's abstract and Specific Aims.) Cystic Fibrosis (CF) is the most common fatal inherited disease in Caucasians. A characteristic pathologic feature is the blockage of exocrine glands in mucus-rich respiratory, gastrointestinal and reproductive tracts which is the basis for the generally held assumption that CF mucus is abnormal. Although the CF gene and gene product, the Cystic Fibrosis transmembrane conductance regulatory (CFTR) have been identified, their relationship to the mucus abnormality is not defined. Since mucin glycoproteins (mucins) are the major macromolecular components of mucus and are responsible for its viscoelastic properties in vivo, alterations in mucins may effect the physiologic behavior of mucus. There are at least four human mucin protein genes expressed in respiratory epithelium; the relative expression of these genes is not known in CF or non-CF epithelium. This application proposes to determine the relative expression of mucin genes in CF and normal epithelium. The long term objective is to elucidate the role of airway mucins in health and disease. The Specific Aims of the present application are: 1) to measure levels of mucin mRNA transcripts in freshly obtained CF and normal nasal epithelial cells and immortalized CF cell lines and matched CF cell lines which have been transfected with the wild-type CFTR gene using a newly developed quantitative mucin RNA method; 2) to test inflammatory mediators known to affect secretion of high molecular weight glycoproteins and known to be present in CF airways to identify agents which modulate expression of specific mucin genes at the transcriptional level in epithelial cells; and 3) to quantitate levels of secreted and cellular mucin proteins in a cell line under resting or stimulated conditions using enzyme linked immunosorbent assays (ELISAs) with specific anti-mucin-peptide antibodies to elucidate whether mucin production is regulated at the transcriptional, translational and/or secretory levels. The information derived from the proposed experiments may fill a major gap in understanding the mucus/mucin abnormality in CF as well as provide basic information about mucin expression in respiratory epithelium.

描述：（根据申请人的摘要和特定目的改编。） 囊性纤维化（CF）是最常见的致命遗传疾病 高加索人。一个特征性的病理特征是阻塞 富含粘液的呼吸道，胃肠道和 生殖道是普遍持有假设的基础 该CF粘液异常。 尽管CF基因和基因产物，但 囊性纤维化跨膜电导调节（CFTR）已 确定，他们与粘液异常的关系尚未定义。 由于粘蛋白糖蛋白（粘蛋白）是主要的大分子 粘液的成分，并负责其粘弹性特性 在体内，粘蛋白的改变可能会影响 粘液。 至少有四个人粘蛋白蛋白基因 呼吸上皮；这些基因的相对表达不是 在CF或非CF上皮中已知。 本申请建议确定 粘蛋白基因在CF和正常上皮的相对表达。 这 长期目标是阐明气道粘蛋白在健康中的作用 和疾病。本应用的具体目的是：1） 在新鲜获得的CF和 正常的鼻上皮细胞和永生的CF细胞系，并匹配 用野生型CFTR基因转染的CF细胞系 使用新开发的定量粘蛋白RNA方法； 2）测试 已知会影响高分子量分泌的炎症介质 糖蛋白并已知存在于CF气道中以识别剂 哪个调节特异性粘蛋白基因在转录上的表达 上皮细胞的水平； 3）定量分泌水平和 在休息或刺激下细胞系中的细胞粘蛋白蛋白 使用酶连接的免疫吸附测定法（ELISA）的条件与 特定的抗粘蛋白肽抗体，以阐明粘蛋白是否 在转录，翻译和/或 分泌级别。从提出的实验得出的信息 可能会填补理解CF中粘液/粘蛋白异常的主要空白 以及提供有关粘蛋白表达的基本信息 呼吸上皮。