ACUTE ISCHEMIC RENAL FAILURE AND KIDNEY-LIVER CROSSTALK

急性缺血性肾衰竭和肾-肝串扰

• 批准号: 6524113
• 负责人: MARIUSZ KIELAR
• 金额: $ 12.93万
• 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/6524113
• 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(改编自应用程序)：从缺血性肾损伤中恢复 受多种细胞因子和生长因子的影响，包括白介素10 IL-10、肿瘤坏死因子(TNFa)、肝细胞生长因子(HGF)。 此前的研究主要集中在肾脏产生这些分子。 然而，在严重的缺血后，肾细胞可能会受到太大的破坏而不能使 调节修复所需的分子。我们认为这样的分子可能是 在肝脏的肾脏之外产生的。拟议的研究将测试 存在调节肾缺血反应的肾-肝轴。 我们认为，缺血肾脏产生IL-6，从而刺激肝脏 产生IL-10并诱导肾脏表达IL-10受体(IL-10R) 使肾脏对IL-10做出反应。这一假设得到了我们的 初步数据显示，肾脏缺血导致肾脏产生 IL-6的表达、IL-10R的表达以及肝脏产生IL-10。 其他研究表明，肝脏IL-10是由肝脏巨噬细胞产生的；我们 发现IL-6刺激巨噬细胞产生IL-10。IL-10已被证明 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是否是来自缺血肾脏的刺激 肝脏产生IL-10；2)检测IL-6是否对 低氧或其他细胞因子，如TNFa和IL-1b；3)检测IL-6的产生 在体外缺氧的肾小管上皮细胞中；4)确定可能的作用 低氧反应元件(NF-IL-6)；5)定位IL-6的产生部位 6)评价核因子-IL-6反应元件是否激活IL-6 并使用基因治疗载体驱动IL-10 在肾脏中的表达。特定目标2将：1)测定IL-10蛋白 肝脏和血液中的存在，然后2)确定IL-10是否抑制 四个已知加剧肾脏缺血的基因(TNFU、ICAM-1、IL-8和iNOS) 3a)研究IL-10对SOCS-3(细胞因子抑制物)的影响 比较SOCS-3在缺血肾组织中的表达 野生型缺血鼠肾组织中IL-10基因敲除及抗IL-10基因表达 抗体处理的小鼠；以及3b)确定SOCS-3在肾脏中是否过度表达 体外细胞抑制肿瘤坏死因子α、细胞间黏附分子-1、白细胞介素8和诱导型一氧化氮合酶的产生 缺氧-复氧。 这项新颖的提议是Kielar博士学习新知识的极好工具 技术，包括器官特异性基因治疗和体内报告基因 化验。培训补助金的发起人陆兆禧博士是一位权威 肾脏免疫学和炎症。共同赞助人罗伯特·芒福德博士是一位 生理反应基因疗法方面的专家。培训计划将 包括免疫学和免疫遗传学的正式课程作业，以准备 申请从事调查工作。