ACUTE ISCHEMIC RENAL FAILURE AND KIDNEY-LIVER CROSSTALK

急性缺血性肾衰竭和肾-肝串扰

• 批准号: 6380143
• 负责人: MARIUSZ KIELAR
• 金额: $ 12.09万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 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 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6380143
• 关键词: acute renal failure; biological signal transduction; cell adhesion molecules; cytokine receptors; genetic regulation; genetically modified animals; interleukin 1; interleukin 10; interleukin 6; interleukin 8; laboratory mouse; laboratory rat; liver; nitric oxide synthase; protein biosynthesis; renal ischemia /hypoxia; tumor necrosis factor alpha

DESCRIPTION (adapted from the application): Recovery from ischemic renal injury is affected by several cytokines and growth factors including interleukin 10 (IL-10), tumor necrosis factor (TNFa), and hepatocyte growth factor (HGF). Previous studies have focused on production of these molecules by the kidney. However, after severe ischemia, the renal cells may be too damaged to make the molecules necessary to regulate repair. We suggest that such molecules may be produced outside the kidney in the liver. The proposed studies will test the existence of a kidney-liver axis that regulates the response to renal ischemia. We propose that the ischemic kidney produces IL-6, which stimulates hepatic production of IL-10 and induces renal expression of the IL-10 receptor (IL-10R) enabling the kidney to respond to IL-10. This hypothesis is supported by our Preliminary Data, demonstrating that renal ischemia results in renal production of IL-6, followed by expression of IL-10R, and hepatic production of IL-10. Others have shown that hepatic IL-10 is produced by hepatic macrophages; we found that IL-6 stimulates macrophages to produce IL-10. IL-10 has been shown to ameliorate ischemic injury to kidney and other organs. Specific Aim 1 will: 1) determine if IL-6 is the signal from the ischemic kidney that stimulates the liver to produce IL-10; 2) test whether IL-6 is produced in response to hypoxia, or to other cytokines, such as TNFa and IL-1b; 3) test IL-6 production in hypoxic renal tubular cells in vitro; 4) determine the role of the putative hypoxia response element (NF-IL-6); 5) localize sites of IL-6 production in the kidney; and 6) evaluate whether the NF-IL-6 response element activates IL-6 gene in the ischemic kidney and use a gene therapy vector to drive IL-10 expression in the kidney. Specific Aim 2 will: 1) determine IL-10 protein presence in the liver and blood and then 2) determine whether IL-10 inhibits four genes (TNFU, ICAM-1, IL-8 and iNOS) known to exacerbate ischemic renal injury; 3a) investigate IL-10 effects on SOCS-3 (suppressor of cytokine signaling-3) upregulation in the ischemic kidney by comparing SOCS-3 expression in the ischemic kidney of wild-type, IL-10 knockout and anti-IL-10 antibody-treated mice; and 3b) determine if over expression of SOCS-3 in renal cells in vitro inhibits production of TNFa, ICAM-1,IL-8 and iNOS during hypoxia-reoxygenation. This novel proposal is an excellent vehicle for Dr. Kielar to learn new techniques, including organ specific gene therapy and in vivo reporter gene assay. The sponsor for the training grant, Dr. Chris Lu, is an authority on renal immunology and inflammation. The co-sponsor, Dr. Robert Munford, is an expert in physiologically responsive gene therapy. The training program will include formal course work in immunology and immunogenetics to prepare the applicant for an investigative career.

描述（改编自本申请）：缺血性肾损伤恢复 受几种细胞因子和生长因子的影响，包括白细胞介素10 （IL-10）、肿瘤坏死因子（TNF α）和肝细胞生长因子（HGF）。 以前的研究集中在肾脏产生这些分子。 然而，在严重缺血后，肾细胞可能受到太大的损伤而不能使肾功能恢复正常。 调节修复所必需的分子。我们认为这些分子可能是 产生于肾脏外的肝脏。拟议的研究将测试 存在调节肾缺血反应的肝肾轴。 我们认为缺血性肾脏产生IL-6，刺激肝脏 产生IL-10并诱导IL-10受体（IL-10 R）的肾表达 使肾脏对IL-10产生反应。这一假设得到了我们的支持。 初步数据，表明肾缺血导致肾生成 IL-6的表达，随后是IL-10 R的表达和IL-10的肝脏产生。 其他研究表明，肝IL-10是由肝巨噬细胞产生的; 发现IL-6刺激巨噬细胞产生IL-10。IL-10已被证实 改善肾脏和其他器官的缺血性损伤。具体目标1将： 1)确定IL-6是否是来自缺血性肾脏的信号， 肝脏产生IL-10; 2）测试IL-6是否响应于 缺氧，或其它细胞因子，如TNF α和IL-1b; 3）测试IL-6的产生 在体外缺氧肾小管细胞中; 4）确定推定的 缺氧反应元件（NF-IL-6）; 5）定位IL-6产生的位点， 肾;和6）评估NF-IL-6应答元件是否激活IL-6 基因，并使用基因治疗载体来驱动IL-10 在肾脏中的表达。具体目标2：1）测定IL-10蛋白 然后2）确定IL-10是否抑制 四种基因（TNFU、ICAM-1、IL-8和iNOS）已知可加重缺血性肾损伤 损伤; 3a）研究IL-10对SOCS-3（细胞因子抑制因子）作用 通过比较SOCS-3表达， 在野生型、IL-10敲除和抗IL-10的缺血肾中， 3b）确定SOCS-3在肾组织中的过表达是否与SOCS-3在肾组织中的过表达有关; 细胞在体外抑制TNF α、ICAM-1、IL-8和iNOS的产生， 缺氧复氧 这一新颖的建议是一个很好的工具，为博士Kielar学习新的 技术，包括器官特异性基因治疗和体内报告基因 比色法培训补助金的赞助商克里斯·卢博士是 肾免疫和炎症。共同发起人罗伯特·芒福德博士是一位 生理反应基因治疗专家培训计划将 包括免疫学和免疫遗传学的正式课程， 申请从事调查工作