Microvascular Endothelial Cells in the Pathogenesis of Chronic Kidney Injury

微血管内皮细胞在慢性肾损伤发病机制中的作用

• 批准号: 7664399
• 负责人: Ikuyo Yamaguchi
• 金额: $ 14.94万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7664399
• 关键词: Adherens Junction; Affect; American; Angiogenic Factor; Animals; Binding; Blood capillaries; Capillary Permeability; Cell Adhesion Molecules; Cell Culture Techniques; Chronic; Chronic Kidney Failure; Co-Immunoprecipitations; Collagen; Complex; Data; Diagnosis; Dialysis procedure; E-Cadherin; Edema; Electron Microscopy; Endocytosis; Endothelial Cells; Explosion; Extravasation; Fibronectins; Fibrosis; Goals; Gold; Heterozygote; Inflammation; Inflammation Mediators; Inflammatory; Injury; Kidney; Kidney Diseases; Kidney Transplantation; Life; Measures; Mediating; Messenger RNA; Methods; Microvascular Permeability; Modeling; Molecular; Mus; Myofibroblast; Pathogenesis; Pathway interactions; Permeability; Phase; Phosphorylation; Placental Growth Factor; Process; Proteins; Public Health; Regulation; Relative (related person); Research; Research Personnel; Role; Severities; Signaling Molecule; Small Interfering RNA; Smooth Muscle Actin Staining Method; Testing; Up-Regulation; Ureteral obstruction; Vascular Endothelial Growth Factors; Vascular Permeabilities; alternative treatment; cadherin 5; capillary; end stage disease; in vitro Model; in vivo; interstitial; knock-down; member; plakoglobin; programs; protein complex; sham surgery; vasoactive agent

DESCRIPTION (provided by applicant): My long-term objective is to understand how renal peritubular capillaries contribute to renal fibrosis, the final common pathway of all chronic kidney diseases. The proposed studies will investigate mechanisms that control peritubular capillary permeability, a major component of the inflammatory process leading to fibrosis, focusing on the adherens junction protein vascular endothelial cadherin (VE-cadherin) and its regulation by the angiogenic signaling molecule placental growth factor (PIGF). Preliminary data show that levels of VE-cadherin and PIGF are increased in the unilateral ureteral obstruction (UUO) model of chronic renal injury. Up-regulation of VE-cadherin may control fibrosis, whereas PIGF appears to be pro-fibrotic. The following aims will investigate how the expression and function of VE-cadherin change during chronic renal injury, test the hypothesis that altering VE-cadherin expression can modulate renal microvascular permeability and fibrosis, and determine whether PIGF acts by altering the function of VE-cadherin: (1) Delineate the changes in renal VE-cadherin expression, localization, and molecular interactions during chronic renal injury. UUO studies will investigate the changes in expression and cellular and subcellular localization of VE-cadherin by immunostaining and immuno-gold electron microscopy, and will also measure VE-cadherin phosphorylation and binding to associated proteins including catenins. (2) Test the hypothesis that up-regulation of VE- cadherin reduces the severity of renal fibrosis by limiting renal microvascular permeability and interstitial inflammation. UUO studies will use VE-cadherin heterozygote () mice and small interfering RNA (siRNA) to investigate how the level of VE-cadherin expression affects the progression of chronic renal injury. (3) Investigate the hypothesis that PIGF induces changes in VE-cadherin function that increase microvascular permeability and promote fibrosis. UUO studies using PIGF -/- mice will determine whether changes in VE- cadherin localization or phosphorylation are associated with modulation of renal fibrosis by PIGF. Cell culture studies will test whether PIGF directly affects VE-cadherin expression and function. This research has a large potential impact on public health, because one in nine Americans currently have chronic kidney disease, and 500,000 have end-stage disease and are on life-long therapy by dialysis or renal transplantation. Thus, alternative treatments for chronic kidney disease are urgently needed.

描述（由申请人提供）： 我的长期目标是了解肾小管周围毛细血管如何导致肾纤维化，这是所有慢性肾脏疾病的最终共同途径。拟议的研究将调查控制管周毛细血管通透性的机制，这是导致纤维化的炎症过程的主要组成部分，重点关注粘附连接蛋白血管内皮钙粘蛋白（VE-钙粘蛋白）及其通过血管生成信号分子胎盘生长因子（PIGF）的调节。初步数据显示，慢性肾损伤的单侧输尿管梗阻（UUO）模型中 VE-钙粘蛋白和 PIGF 水平升高。 VE-钙粘蛋白的上调可以控制纤维化，而 PIGF 似乎是促纤维化的。以下目的将研究慢性肾损伤期间 VE-cadherin 的表达和功能如何变化，检验改变 VE-cadherin 表达可以调节肾脏微血管通透性和纤维化的假设，并确定 PIGF 是否通过改变 VE-cadherin 的功能发挥作用：（1）描述慢性肾损伤期间肾脏 VE-cadherin 表达、定位和分子相互作用的变化。 UUO 研究将通过免疫染色和免疫金电子显微镜研究 VE-钙粘蛋白的表达以及细胞和亚细胞定位的变化，还将测量 VE-钙粘蛋白的磷酸化以及与包括连环蛋白在内的相关蛋白的结合。 (2)检验VE-钙粘蛋白上调通过限制肾微血管通透性和间质炎症来降低肾纤维化严重程度的假设。 UUO研究将使用VE-钙粘蛋白杂合子（）小鼠和小干扰RNA（siRNA）来研究VE-钙粘蛋白表达水平如何影响慢性肾损伤的进展。 (3) 研究 PIGF 诱导 VE-钙粘蛋白功能变化从而增加微血管通透性并促进纤维化的假设。使用 PIGF -/- 小鼠进行的 UUO 研究将确定 VE-钙粘蛋白定位或磷酸化的变化是否与 PIGF 对肾纤维化的调节有关。细胞培养研究将测试 PIGF 是否直接影响 VE-钙粘蛋白的表达和功能。 这项研究对公众健康具有巨大的潜在影响，因为目前有九分之一的美国人患有慢性肾病，50万人患有终末期疾病，并通过透析或肾移植进行终身治疗。因此，迫切需要慢性肾病的替代治疗。