RENIN EXPRESSING CELL LINES FOR HYPERTENSION RESEARCH

用于高血压研究的肾素表达细胞系

• 批准号: 2224512
• 负责人: KENNETH W GROSS
• 金额: $ 25.71万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 1996-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2224512
• 关键词: DNA footprinting; SDS polyacrylamide gel electrophoresis; cathepsin B; cell line; cell type; clone cells; gene expression; genetic regulatory element; genetic transcription; genetically modified animals; immunoprecipitation; in situ hybridization; kidney cell; laboratory mouse; molecular cloning; neoplastic cell culture for noncancer research; nucleic acid sequence; oncogenes; polymerase chain reaction; posttranslational modifications; protein biosynthesis; protein sequence; protein structure; radiotracer; renin; secretion; subtraction hybridization; transfection; tumor antigens; western blottings

Development of a detailed understanding of the regulation of renin gene expression, biosynthesis and secretion has been hampered by the lack of suitable cell culture models for the juxtaglomerular (JG) cell - the classical renal site of active renin release into the circulation. We have utilized the approach of transgene targeted oncogenesis to induce neoplasia in renin expressing cells of renal origin and have isolated and established a clonal cell line in in vitro culture. Preliminary studies suggest that this cell line has retained significant features characteristic of the differentiated state of its in vivo counterpart and thus should provide a unique resource with which to investigate important issues of renin gene expression. Most significantly, the 4.1 cell line exhibits high level expression of the endogenous renin gene (Ren 1c) as manifest in abundant renin. Moreover, exogenous renin genes introduced by transfection also exhibit high level expression. We thus propose to investigate the utility of these cells as an assay to define in detail the cis-acting DNA sequences pertinent to renal renin gene expression, and the transcriptional machinery which serves to give the JG cell its unique identity. We will also examine the utility of this cell line to assess mechanisms, and identify components, relevant to the targeting, processing and secretion of the renin polypeptide. It should be invaluable to assess and compare the results obtained with this cell line, which expresses its endogenous renin gene and is thought to be derived from a naturally occurring site of active renin generation, with those previously obtained from heterologous transfected cell systems. Finally, we propose to expand upon our current base to develop renal cell lines harboring conditional oncogenes and to develop representative cell lines from extra-renal sites. Cell lines obtained by conditional oncogenesis may afford opportunities for retention of increased differentiative character and thus provide refined tools for further analysis. Cell lines derived from other tissue sites or organ sites will provide important tools for more detailed analysis of the molecular basis for differential tissue specific expression of the murine renin genes.

对肾素基因调节的详细理解的发展 缺乏表达，生物合成和分泌受到阻碍 近齿状细胞（JG）细胞的合适细胞培养模型 - 活动肾素的经典肾脏位点释放到循环中。 我们有 利用转基因靶向肿瘤发生的方法诱导肿瘤 在表达肾脏起源细胞的肾素中，已分离和建立 体外培养中的克隆细胞系。 初步研究表明 该单元线保留了重要特征的特征 其体内差异化状态，因此应提供 研究肾素基因重要问题的独特资源 表达。 最重要的是，4.1单元线表现出高水平 内源性肾素基因（REN 1C）的表达在丰富的 肾素。 此外，也通过转染引入的外源肾素基因 表现出高水平的表达。 因此，我们建议调查效用 这些细胞作为详细定义顺式作用DNA序列的测定法 与肾素肾素基因的表达和转录机械有关 这使JG单元具有其独特的身份。 我们还将检查 该细胞系的实用性评估机制，并确定 组件，与针对，处理和分泌有关的组件 肾素多肽。 评估和比较 用该细胞系获得的结果，该细胞系表示其内源性肾素 基因，被认为是从天然存在的活性位点得出的 肾素产生，先前从异源获得的那些 转染的细胞系统。 最后，我们建议扩展我们的当前 发展有条件性癌基因的肾细胞系和 从肾外部位发展代表性的细胞系。 细胞系 通过有条件的肿瘤形成获得的可能有机会保留的机会 具有较高的差异化特征，从而为 进一步分析。 衍生自其他组织部位或器官的细胞系 网站将提供重要的工具，以更详细地分析 鼠的分子基础鼠的特异性表达 肾素基因。