Role of Nlrp3 in Ischemic Organ Injury

描述(由申请人提供)：这项建议研究了一个关键的先天免疫受体Nlrp3如何在小鼠肾缺血再灌注(IR)损伤模型中对肾小管上皮(RTE)细胞损伤起作用。该项目对临床所致的缺血性肾损伤(如供肾移植)具有重要意义。广泛/长期目标：拟议研究的长期目标是确定Nlrp3如何促进肾脏的损伤组织反应。具体目的：本提案的具体目的是验证细胞质PRR Nlrp3是肾IR损伤所致RTE细胞损伤的关键因素这一假设。目的1探讨Nlrp3激活是否直接导致RTE细胞损伤，确定导致这种损伤的信号事件，并确定Nlrp3阻断是否能阻止RTE细胞损伤。目的2探讨Nrp3的激活是否主要通过直接(局部)或间接(全身)机制参与肾脏IR损伤。研究设计和实现上述目标的方法：目的1将测试Nlrp3连接如何损伤RTE细胞，Nlrp3激活刺激是否指导RTE细胞损伤的模式，从坏死细胞(DAMP)释放的分子如何激活Nlrp3介导的健康RTE细胞损伤，以及是否可以通过阻断Nlrp3或其上游激活途径之一来阻断RTE细胞损伤。在体内，IR损伤合并了其他组织损伤的介质，如炎症，因此目的2关注Nlrp3的激活是否通过分离局部(肾脏损伤)和全身(炎症)在肾脏中发挥更广泛的作用。在肾移植模型中，直接(局部的，肾脏特异性的)效应与间接(全身)效应是分开的，其中在野生型(WT)宿主中研究Nlrp3/-移植肾的损伤反应，在Nlrp3-/-宿主中研究WT肾损伤。项目的健康相关性：如果这项提案的目标得以实现，我们将了解受损组织释放的分子如何激活肾脏中依赖Nlrp3的损伤反应。这一知识对于开发合理的靶向疗法以预防或改善预期缺氧的临床情况下的肾脏IR损伤(例如，在供肾移植前对供体肾脏进行预处理)至关重要。专注于缺血性肾损伤的早期事件是有效治疗策略的最大希望。

DESCRIPTION (provided by applicant): This proposal studies how a key innate immune receptor, Nlrp3, contributes to renal tubular epithelial (RTE) cell injury in a murine model of renal ischemia reperfusion (IR) injury. The project is highly significant for clinical obligate ischemic kidney injury (e.g., as in donor kidneys harvested for transplantation). Broad/long-term objectives: The long-term goals of the proposed research are to define how Nlrp3 contributes to injurious tissue responses in the kidney. Specific Aims: The specific objective of this proposal is to test the hypothesis that the cytoplasmic PRR Nlrp3 is a key contributor to RTE cell damage induced by renal IR injury. Aim 1 asks whether Nlrp3 activation directs RTE cell injury, defines the signaling events that lead to this injury, and determines whether Nlrp3 blockade prevents RTE cell injury. Aim 2 asks whether activation of Nrp3 contributes to renal IR injury primarily through direct (local) or indirect (systemic) mechanisms. Research Design and Methods for Achieving the Stated Goals: Aim 1 will test how ligation of Nlrp3 injures RTE cells, whether the Nlrp3 activating stimulus directs the mode of RTE cell injury, how molecules released from necrotic cells (DAMPs) activate Nlrp3-mediated injury of healthy RTE cells, and whether RTE cell injury can be blocked by either blocking Nlrp3 or one of its upstream activating pathways. In vivo IR injury incorporates other mediators of tissue injury, such as inflammation, so aim 2 focus on whether Nlrp3 activation plays a broader role in the kidney by separating local (kidney injury) from systemic (inflammation). Direct (local, kidney specific) effects are separated from indirect (systemic) effects in a kidney transplant model where injury responses of the Nlrp3-/- transplanted kidney are studied in a wild type (WT) host, and WT kidney injury studied in a Nlrp3-/- host. Health Relatedness of Project: If the aims of this proposal are met we will learn how molecules released from injured tissue activate Nlrp3-dependent injurious responses in the kidney. This knowledge is crucial for the development of rational target therapies for prevention or amelioration of renal IR injury in clinical situations where hypoxia is anticipated (e.g., pretreatment of donor kidneys prior to harvest for transplantation). Focusing on the earliest events of ischemic kidney injury holds the greatest promise for effective therapeutic strategies.