ROLE OF CELL CYCLE PROTEINS IN GLOMERULAR EPITHELIAL CELL GROWTH

细胞周期蛋白在肾小球上皮细胞生长中的作用

• 批准号: 6357051
• 负责人: Stuart James Shankland
• 金额: $ 12.07万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6357051
• 关键词: Wilms' tumor; cell growth regulation; cell proliferation; cellular pathology; cyclin dependent kinase; cyclins; enzyme inhibitors; flow cytometry; gene expression; gene targeting; genetically modified animals; glomerulonephritis; glomerulosclerosis; histogenesis; in situ hybridization; kidney cell; kidney disorder; kidney hypertrophy; laboratory mouse; laboratory rat; northern blottings; renal glomerulus; tissue /cell culture; transcription factor; western blottings

Description: (Taken directly from the application) The glomerular visceral epithelial cell (VEC) or podocyte is injured in immune and non-immune mediated glomerular disease. In contrast to the marked proliferation that follows injury to the glomerular mesangial and endothelial cells, VEC do not proliferate but rather hypertrophy. These growth responses of VEC are maladaptive and underlie the development of progressive glomerulosclerosis. Cell growth is controlled at the level of the cell cycle by cell cycle regulatory proteins. Proliferation requires that cyclin-dependent kinases (CDK) be activated by specific cyclins. Cyclin kinase inhibitors (CKI) inhibitor proliferation by inhibiting cyclin-CDK complexes, but a role for CKI in mediating cell hypertrophy is not known. This project will test the hypothesis that specific CKIs (p21, p27 and p57) determine the VEC growth response to injury. In the first specific aim, we determine the VEC growth response to injury. In the first specific aim, we propose that VEC can proliferate in vitro and in experimental models of glomerular disease in the levels of CKI are lowered, and that if CKI levels are increased following injury, VEC will not proliferate, but rather hypertrophy. To lower CKI levels in vitro, we will use VEC in culture derived from CKI (p21, p27 and p57) knockout nd wild-type mice. VEC in vitro will be stably transfected to increase CKI levels. A role for CKI in determining the growth response (proliferation, hypertrophy) in vivo will be determined by inducing immune and non-immune forms of VEC injury in CKI (p21, p27 and p57) knockout and wild-type mice. In the second aim, we will test the hypothesis that the Wilms Tumor gene product (WT-1), a transcriptional activator or suppressor and a protein required for normal kidney development, inhibits VEC proliferation and that this is due to the up-regulation of specific CD1. Accordingly, we will measure proliferation in VEC in vitro where the levels of WT-1 are increased (Over-expression) or decreased (antisense), and correlate this with the levels of CKI (p21, p27 and p57). The expression of WT-1 and CKI will be correlated in a variety of animal models of glomerular injury to VEC, and during normal glomerulogenesis. The data generated will provide novel insights into the events that precede and underlie progressive glomerulosclerosis, and should lead to opportunities for potential new therapeutic interventions.

描述：（直接取自应用程序）肾小球内脏 上皮细胞（VEC）或足细胞在免疫和非免疫中受到损伤 介导的肾小球疾病。相比之下， 在肾小球系膜细胞和内皮细胞损伤后， 不是增殖而是肥大。VEC的这些生长反应是 适应不良，是渐进式发展的基础 肾小球硬化 细胞生长是在细胞周期水平上被细胞周期控制的 调节蛋白增殖需要细胞周期蛋白依赖性激酶 (CDK)被特定的细胞周期蛋白激活。细胞周期蛋白激酶抑制剂（CKI） 通过抑制细胞周期蛋白-CDK复合物来抑制增殖，但对于 尚不清楚CKI介导细胞肥大。 本项目将检验特定CKI（p21，p27和p27） p57）确定VEC对损伤的生长反应。在第一个具体的 目的是确定血管内皮细胞对损伤的生长反应。上 具体而言，我们提出血管内皮细胞可以在体外和 肾小球疾病的CKI水平的实验模型是 降低，如果CKI水平在受伤后增加，VEC将 不是增殖而是肥大为了降低体外CKI水平，我们 将使用来源于CKI（p21，p27和p57）敲除的培养物中的VEC， 野生型小鼠。VEC在体外将被稳定转染以增加CKI 程度. CKI在决定生长反应（增殖， 将通过诱导免疫和非免疫细胞（例如，细胞肥大）在体内的表达来确定。 CKI（p21、p27和p57）敲除和野生型小鼠中VEC损伤的形式。 在第二个目标中，我们将测试Wilms肿瘤基因 产物（WT-1）、转录激活因子或抑制因子和蛋白质 肾脏正常发育所需，抑制VEC增殖， 这是由于特异性CD 1的上调。因此我们 将在体外测量VEC的增殖，其中WT-1的水平是 增加（过度表达）或减少（反义），并将其与 CKI（p21、p27、p57）表达水平的变化。WT-1和CKI的表达 在各种肾小球损伤的动物模型中， VEC，并在正常肾小球形成。生成的数据将提供 对发生在进步主义之前和之下的事件的新见解 肾小球硬化症，并应导致潜在的新的机会， 治疗干预。