Understanding the Function of Tamm-Horsfall Protein in Acute Kidney Injury

急性肾损伤(AKI)是住院退伍军人死亡率和发病率的主要决定因素。 了解AKI的病理生理学对于治疗的发展至关重要。此应用程序 建议研究Tamm-Horsfall蛋白(THP)在AKI中的作用。这种蛋白质只在细胞中表达 肾的粗大升支(TAL)的细胞。在上一个资金周期中，我们发现了一个关键角色 THP在AKI中调节保护性小管串扰中的作用。我们发现THP可以调节炎症 S3段的信号和损伤，S3段是肾脏外髓中与TAL相邻的小管。这个 目前提案的直接目标是研究THP与S3段在 调节急性心肌梗死患者中性粒细胞的渗透。这些研究将形成过渡到治疗的基础 申请。事实上，PI的长期目标是了解THP的作用如何被调节以治疗 发展AKI的退伍军人。 中心假说是Tamm-Horsfall蛋白调节由 外髓的缺血性损伤；一种需要THP与周围小管相互作用的效应 元素。这一假设是建立在强有力的初步数据和已公布数据的基础上的。这个 以下具体目标将被用来研究这一假设。 目的1探讨THP在急性肾损伤后肾组织中性粒细胞浸润中的重要调节作用。 目的2将研究THP在肾脏过程中对邻近上皮细胞的特定炎症通路的调节 伤害 目的2研究THP对急性心肌梗死后抑制中性粒细胞内流和促进恢复的治疗作用 这项研究将使用THP基因敲除和野生型小鼠。AKI使用的动物模型是肾脏 缺血再灌注损伤可通过夹闭肾蒂实现。这项工作中使用的其他技术 包括：特定管状切片的免疫荧光激光显微解剖、免疫荧光共聚焦 活体显微镜，流式细胞仪，实时聚合酶链式反应，蛋白质印迹，各种形式的层析和 质谱学。 这项研究是创新的，因为它将揭示THP的新调节功能，这些功能可能与 不仅在肾脏损伤中，而且在其他形式的急性或慢性肾病中也是如此。未来战略将增强 THP在肾脏中的作用将具有重要的治疗意义。

Acute kidney injury (AKI) is a major determinant of mortality and morbidity in hospitalized veterans. Understanding the pathophysiology of AKI is essential for the development of therapy. This application proposes to study the role of Tamm-Horsfall protein (THP) in AKI. This protein is expressed exclusively in the kidney by cells of the thick ascending limbs (TAL) of Henle. In the last funding cycle, we uncovered a key role of THP in mediating a protective tubular cross-talk in AKI. We showed that THP regulates inflammatory signaling and injury to S3 segments, which are neighboring tubules to TAL in the kidney outer medulla. The immediate goal of the current proposal is to study the importance of the interaction of THP with S3 segments in regulating neutrophil infiltration in AKI. These studies will form the basis to transition into therapeutic applications. Indeed, the PI's long term goal is to understand how the role of THP can be modulated to treat veterans who develop AKI. The central hypothesis is that Tamm-Horsfall protein regulates the inflammatory response triggered by ischemic injury in the outer medulla; an effect that requires an interaction of THP with the surrounding tubular elements. This hypothesis has been formulated on the basis of strong preliminary and published data. The following specific aims will be used to investigate this hypothesis. Aim 1 will investigate the role of THP as a key regulator of neutrophil infiltration in the kidney after AKI. Aim 2 will study specific inflammatory pathways regulated by THP in the neighboring epithelium during kidney injury Aim 2 will investigate therapeutic modulation of THP to inhibit neutrophil influx and promote recovery after AKI This research will involve the use of THP knockout and wild type mice. The animal model used for AKI is renal ischemia-reperfusion injury achieved through renal pedicle clamping. Other techniques used in this work include: Immunofluorescence laser micro-dissection of specific tubular sections, Immunofluoresence confocal and intravital microscopy, flow cytometry, real time-PCR, western blot, various forms of chromatography and mass spectrometry. This research is innovative, because it will uncover novel regulatory functions for THP that may be relevant not only in kidney injury but also in other forms of acute or chronic renal disease. Future strategies that enhance the role of THP in the kidney will be of important therapeutic significance.