The Role of Caspases in Warm and Cold Ischemia

Caspases 在热缺血和冷缺血中的作用

基本信息

批准号：
7918748
负责人：
Alkesh Harihar Jani
金额：
$ 13.11万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-15 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7918748
关键词：
Acute Acute Kidney Failure Allografting Antigens Apoptosis Apoptotic Applications Grants Brush Border Caspase Caspase Inhibitor Caspase-1 Cell Death Cells Cessation of life Chronic Chronic rejection of renal transplant Cryopreservation Data Development Epithelial Cells Functional disorder Gene Expression Grant Immune Sera Immunohistochemistry Incidence Inflammatory Injury Interferon Type II Interferons Interleukin-18 Ischemia Kidney Kidney Diseases Kidney Transplantation MHC Class I Genes Mediating Mediator of activation protein Messenger RNA Mitochondria Modeling Morphology Mus Necrosis Organ Organ Preservation Organ Preservation Solutions Perfusion Production Proteins Protocols documentation Publishing Reperfusion Therapy Research Personnel Role Transplantation Tubular formation Warm Ischemia base caspase-3 caspase-8 caspase-9 clinically relevant cytochrome c cytokine delayed graft function immunogenicity implantation inhibitor/antagonist interferon-alpha B interstitial mRNA Expression mouse development prevent programs receptor research study tubular necrosis

项目摘要

Delayed graft function (DGF) independently predicts reduced 1- and 5- year kidney transplant survival and contributes to the development of chronic allograft nephropathy (CAN). DGF is caused by both cold and warm ischemia. Cold ischemia occurs during both pulsatile machine perfusion and static cold storage of kidneys. This grant will focus on the pathophysiology of cold and warm ischemia and specifically the role of caspases. Caspase-3 is a major mediator of apoptotic and necrotic cell death. Caspase-1 converts the pro- inflammatory cytokines IL-1beta and IL-18 to their active forms. The overall hypothesis is that caspases mediate proximal tubular damage in both cold and warm ischemia. In a model of cold ischemia the contribution of caspase-3 to proximal tubular apoptosis as well as severe brush border injury will be assessed. Gene expression, protein and activity of caspases-3, 7, 9 (mitochondrial pathway of apoptosis) and 8 (death receptor pathway of apoptosis) will be determined during both static and pulsatile cold ischemia. Immunohistochemistry for activated caspase-3 in proximal tubules will be performed. The effect of caspase inhibition on kidney morphology will be assessed. The findings of these experiments will contribute to the understanding of cold ischemia injury and organ preservation. Caspase-1-mediated production of IL-18 mediates ischemic ARF. IL-18 is a potent inducer of interferbn gamma, which is a powerful inducer of MHC expression. Caspase-1 and IL-18-mediated increase in interferon gamma expression in the kidney and MHC expression on proximal tubules will be determined in a model of warm ischemia. Interferon gamma and MHC class I and II mRNA expression in the kidney will be assessed. The effect of caspase-1, IL-18 and interferon gamma inhibition on MHC expression will be evaluated. Reduced MHC expression is associated with decreased kidney graft immunogenicity and has important implications for the deleterious effect of DGF on both short and long-term allograft function.

移植物功能延迟 (DGF) 独立预测肾移植 1 年和 5 年存活率的降低有助于慢性同种异体移植肾病（CAN）的发展。 DGF 是由寒冷和热缺血。脉动机器灌注和静态冷藏过程中都会发生冷缺血肾脏。这笔资金将重点研究冷和热缺血的病理生理学，特别是半胱天冬酶。 Caspase-3 是细胞凋亡和坏死性细胞死亡的主要介质。 Caspase-1 将亲炎症细胞因子 IL-1beta 和 IL-18 恢复其活性形式。总体假设是半胱天冬酶介导冷缺血和热缺血时的近端肾小管损伤。在冷缺血模型中 caspase-3 对近端肾小管凋亡以及严重刷状缘损伤的贡献将是评估。 Caspases-3、7、9（线粒体凋亡途径）的基因表达、蛋白和活性和8（细胞凋亡的死亡受体途径）将在静态和脉冲冷期间测定缺血。将进行近曲小管中活化的 caspase-3 的免疫组织化学分析。效果将评估半胱天冬酶抑制对肾脏形态的影响。这些实验的结果将有助于了解冷缺血损伤和器官保存。 Caspase-1介导的 IL-18 的产生介导缺血性 ARF。 IL-18 是γ干扰素的有效诱导剂，γ干扰素是一种 MHC 表达的强大诱导剂。 Caspase-1 和 IL-18 介导的干扰素 γ 增加肾脏中的表达和近端小管上的 MHC 表达将在温热模型中确定缺血。将评估肾脏中的干扰素 γ 以及 MHC I 类和 II 类 mRNA 表达。这将评估 caspase-1、IL-18 和干扰素 γ 抑制对 MHC 表达的影响。减少 MHC 表达与肾移植免疫原性降低相关，具有重要意义 DGF 对短期和长期同种异体移植功能的有害影响。