IMMORTAL SWEAT GLAND CELLS FOR CYSTIC FIBROSIS RESEARCH

用于囊性纤维化研究的永生汗腺细胞

• 批准号: 3242038
• 负责人: PAUL M QUINTON
• 金额: $ 15.72万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3242038
• 关键词: cystic fibrosis; electrophysiology; electroporation; gel electrophoresis; gene expression; human tissue; membrane permeability; membrane potentials; secretion; sweat glands; tissue /cell culture; transfection

Considerable progress has been made in CF research since the demonstration of low Cl- permeability in epithelia of target organs. Recent studies by others on cultured tracheal epithelial cells indicate that the defect in Cl- transport is not in the Cl- channels, but a malfunction in regulation of the Cl- pathway. Advances at the molecular level have been no less exciting. Linkage studies have localized the CF gene to chromosome 7q22-31 and a candidate for the cystic fibrosis locus has now been isolated. Identification of the CF gene and its product is only a beginning. Definition of the metabolic defect require an understanding of the abnormal gene product and its physiological expression. The goal of the research in the proposed project is to establish immortal cell lines from the human eccrine sweat gland for use as a model system for studying the basic physiological and biochemical defects in CF. The human eccrine sweat gland is an appropriate model for CF research, since sweat glands from CF subjects: i) demonstrate defects in secretory and reabsorptive functions ii) appear to express differences in the phosphorylation of a specific polypeptide iii) are not burdened by other pathological manifestations; and iv) are relatively accessible. Establishing cultured cell lines from the sweat gland will increase the quantity and longevity of the tissue and provide the opportunity to study and compare the two regions of the sweat gland (the secretory coil and reabsorptive duct) independently and in a simplified model. We propose to characterize the cells in culture to determine the extent to which they resemble the progenitor tissue, and more importantly to demonstrate the persistence of the phenotypic expression of the CF genotype in the cultured cells. Specifically, we will perform electrophysiological studies of Cl- conductance its regulation by various agonists as well as analyses of phosphorylation profiles of polypeptides. Expression of the characteristic CF phenotype in culture will allow us to investigate the underlying metabolic defect. Eventually we hope to be able to alter its expression toward a therapeutic control of CF pathology.

自从CF研究中取得了很大进展 证明靶标上皮中的低Cl-渗透性 器官。 其他人的最新研究对培养的气管上皮 细胞表明Cl-转运中的缺陷不在Cl-中 通道，但在调节CL途径中的故障。 分子水平的进步同样令人兴奋。 连锁研究将CF基因定位于7q22-31染色体 囊性纤维化基因座的候选者现已 孤立。 CF基因及其产物的识别仅是 一个开始。 代谢缺陷的定义需要 了解异常基因产物及其生理 表达。 拟议项目中研究的目的是建立 来自人类eccrine汗腺的不朽细胞系用作 研究基本生理和生化的模型系统 CF中的缺陷。 人类eccrine汗腺是合适的 CF研究模型，因为CF受试者的汗腺：i） 表现出分泌物和重吸收功能的缺陷ii） 在特定的磷酸化中似乎表达了差异 多肽III）不受其他病理的负担 表现；和iv）相对易于访问。 建立 汗腺中培养的细胞系将增加数量 和组织的寿命，并提供了学习的机会 并比较汗腺的两个区域（分泌线圈 并在简化的模型中独立地进行了重吸管。 我们建议表征培养细胞以确定 它们类似于祖细胞组织的程度，更多 重要的是要证明表型的持久性 CF基因型在培养细胞中的表达。 具体来说， 我们将对cl-电导的电生理研究 各种激动剂的调节以及分析 多肽的磷酸化谱。 表达 文化中的特征CF表型将使我们能够调查 潜在的代谢缺陷。 最终我们希望能够 改变其表达对CF病理的治疗控制。