Role of Nlrp3 in Ischemic Organ Injury

描述（由申请人提供）：本提案研究关键先天免疫受体Nlrp 3如何在肾缺血再灌注（IR）损伤的鼠模型中促进肾小管上皮（RTE）细胞损伤。该项目对于临床上的专性缺血性肾损伤（例如，如用于移植的供体肾）。广泛/长期目标：拟议研究的长期目标是确定Nlrp 3如何促进肾脏中的损伤组织反应。具体目标：本提案的具体目的是检验细胞质PRR Nlrp 3是肾IR损伤诱导的RTE细胞损伤的关键因素的假设。目的1询问Nlrp 3激活是否指导RTE细胞损伤，定义导致这种损伤的信号传导事件，并确定Nlrp 3阻断是否防止RTE细胞损伤。目的2探讨Nrp 3的激活是否主要通过直接（局部）或间接（全身）机制导致肾IR损伤。研究设计和实现既定目标的方法：目标1将测试Nlrp 3的连接如何损伤RTE细胞，Nlrp 3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Nlrp 3介导的健康RTE细胞损伤，以及RTE细胞损伤是否可以通过阻断Nlrp 3或其上游激活途径之一来阻断。体内IR损伤结合了组织损伤的其他介质，例如炎症，因此目的2关注Nlrp 3活化是否通过将局部（肾损伤）与全身性（炎症）分开而在肾脏中发挥更广泛的作用。在肾移植模型中将直接（局部、肾特异性）作用与间接（全身）作用分开，其中在野生型（WT）宿主中研究Nlrp 3-/-移植肾的损伤应答，并在Nlrp 3-/-宿主中研究WT肾损伤。项目的健康相关性：如果这个提议的目标得以实现，我们将了解从受损组织释放的分子如何激活肾脏中依赖Nlrp 3的损伤反应。这一知识对于开发合理的靶向治疗以预防或改善预期缺氧的临床情况下的肾IR损伤是至关重要的（例如，在收获用于移植之前对供体肾进行预处理）。关注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.