The immunomodulatory 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对急性心肌梗死患者中性粒细胞渗入的调节作用目前的提案通过在基础上扩展这些发现 强有力的初步数据表明，THP通过增强愈合功能在AKI的康复中起着重要作用 指肾单核巨噬细胞(MPC)，如巨噬细胞。这些研究将形成以下基础 转变为治疗应用。事实上，PI的长期目标是了解THP的作用如何 可以被调节来治疗患AKI的退伍军人。 中心假设是Tamm-Horsfall蛋白对于促进AKI的康复至关重要，它通过增强 修复所需的肾脏MPC的丰度和活性。这一假说是在 有强大的初步数据和公布数据的基础。将使用以下具体目标来调查这一点 假设。 目标1将明确THP在急性肾损伤后肾脏MPC内环境稳定中的作用 目标2将确定THP在AKI恢复过程中对肾脏MPC活性的作用 这项研究将使用THP基因敲除和野生型小鼠。一种组织特异性基因敲除小鼠将 也可用于研究THP对MPC信号转导的影响。用于AKI的动物模型是肾缺血- 通过夹闭肾蒂实现再灌注损伤。这项工作中使用的其他技术包括： 免疫荧光共聚焦3D成像和组织细胞术，创新的大规模多路成像， 流式细胞术、实时荧光定量聚合酶链式反应、蛋白质印迹、蛋白质纯化和药理给药。 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 regulating neutrophil infiltration in AKI. The current proposal expands these findings by building on strong preliminary data showing that THP is important in recovery from AKI by enhancing the healing functions of renal mononuclear phagocytic cells (MPCs) such as macrophages. 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 is essential to promoting recovery from AKI by enhancing the abundance and activity of renal MPCs needed for healing. 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 define the role of THP on the homeostasis of renal MPCs after AKI Aim 2 will define the role of THP on the activity of renal MPCs during recovery from AKI This research will involve the use of THP knockout and wild type mice. A tissue specific knockout mouse will also be used to study the effect of THP on MPC signaling. The animal model used for AKI is renal ischemia- reperfusion injury achieved through renal pedicle clamping. Other techniques used in this work include: Immunofluorescence confocal 3D imaging and tissue cytometry, innovative large scale multiplexed imaging, flow cytometry, real time-PCR, western blot, protein purification and pharmacological dosing. 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.