The immunomodulatory function of Tamm-Horsfall protein in acute kidney injury

急性肾损伤（阿基）是住院退伍军人死亡率和发病率的主要决定因素。 了解阿基的病理生理学对于开发治疗至关重要。本申请 提出研究Tamm-Horsfall蛋白（THP）在阿基中的作用。这种蛋白质只表达在 肾由Henle氏粗升支（TAL）细胞构成。在上一个融资周期中，我们发现了一个关键角色， THP对阿基中性粒细胞浸润的调节作用。目前的建议通过建立在 强有力的初步数据表明，THP通过增强愈合功能在阿基的恢复中很重要 肾单核吞噬细胞（MPC），如巨噬细胞。这些研究将构成基础， 过渡到治疗应用。事实上，PI的长期目标是了解THP的作用 可以调节以治疗发生阿基的退伍军人。 中心假设是Tamm-Horsfall蛋白通过增强AKI的免疫应答而对促进AKI恢复至关重要。 愈合所需的肾MPC的丰度和活性。这一假设是根据 根据初步和公布的有力数据。以下具体目标将用于调查这一点 假说. 目的1将明确THP对阿基后肾MPCs稳态的作用 目的2将确定THP在阿基恢复期间对肾MPCs活性的作用 这项研究将涉及使用THP敲除和野生型小鼠。组织特异性敲除小鼠将 也可用于研究THP对MPC信号传导的影响。用于阿基的动物模型是肾缺血- 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，创新的大规模多路复用成像， 流式细胞术、真实的时间-PCR、蛋白质印迹、蛋白质纯化和药理学剂量。 这项研究是创新的，因为它将揭示THP的新的调节功能， 不仅在肾损伤中，而且在其它形式的急性或慢性肾病中。未来的战略， 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.