TIGIT in acute kidney injury and repair

摘要/项目摘要 急性肾损伤(AKI)在自体肾脏和同种异体移植肾中的发生率都很高，并且没有特殊的治疗方法。 可用。先前的研究已经确定T细胞的激活和运输是一种重要的机制， 调节缺血再灌注(IR)和肾毒性AKI，以及其他重叠的免疫和非 免疫机制。此外，AKI在接受免疫检查点抑制剂治疗的患者中很常见。 靶向细胞毒性T淋巴细胞相关抗原4(CTLA4)和程序性细胞死亡受体1(PD1) 多发性癌症。我们使用RNA测序和流式细胞术的初步数据显示显著表达 人T细胞中新型免疫检查点分子--T细胞免疫受体(TIGIT)的研究 缺血再灌注后小鼠肾脏和人缺血肾脏。最近公布的数据表明，TIGIT协同抑制 激活修饰Th1和Th17反应，并调节Treg抑制活性。我们的初步数据显示 小鼠肾脏表达TIGIT的T细胞高度活化并产生促炎细胞因子 IR.重要的是，在IR和顺铂AKI模型中，缺乏TIGIT(TIGIT KO)的小鼠受到AKI的保护。 表明TIGIT在AKI期间扮演了不利的角色。因此，了解TIGIT介导的炎症 AKI期间的反应对于开发新的AKI疗法和减轻AKI的肾脏副作用至关重要 免疫检查点疗法。这一提议的中心假设是TIGIT促进促炎 肾脏T细胞功能及Treg抑制功能受损。为了检验这一假设，我们将(目标1) 体外和体外研究TIGIT对小鼠肾脏T细胞表型、功能和转录的影响 在活体内接近。我们将进一步研究TIGIT和它的共同信号之间的功能关系 调节肾脏T细胞功能的伙伴(CD226、CD155)和其他共抑制分子(PD1、CTLA4) 在基线和AKI期间。此外(目标2)，我们将测试T细胞特异性TIGIT活性是 体内采用过继转移方法驱动AKI并损害修复过程的主要机制 抗TIGIT激动剂/拮抗剂抗体对WT和TIGIT KO小鼠AKI结局的影响及阻断TIGIT AKI建立后修复阶段的信号转导。最后(目标3)，我们将研究TIGIT的功能效应 人肾T细胞在肾癌患者和活体肾活检中的表达我们还将 TIGIT在缺血死亡供肾标本分离的T细胞上的表达 活体供肾T细胞在单个细胞水平上的转录效应。这些研究的结果将是 首先就TIGIT介导的肾脏T细胞功能、治疗潜力提供重要信息 将TIGIT作为AKI治疗的靶点，并为未来的临床前和临床研究奠定基础。

ABSTRACT/PROJECT SUMMARY Acute kidney injury (AKI) occurs at a high rate in both native kidneys and allografts and has no specific therapy available. Prior studies have established T cell activation and trafficking as an important mechanism that modulate ischemia reperfusion (IR) and nephrotoxic AKI, along with other overlapping immune and non- immune mechanisms. Furthermore, AKI is common in patients treated with immune checkpoint inhibitors targeting cytotoxic T lymphocyte-associated antigen 4 (CTLA4) and programmed cell death receptor 1 (PD1) for multiple cancers. Our preliminary data using RNA sequencing and flow cytometry shows significant expression of novel immune checkpoint molecule, T cell immunoreceptor with Ig and ITIM domains (TIGIT) in T cells from post IR mouse kidney and ischemic human kidney. Recently published data suggest that TIGIT co-inhibitory activity modifies Th1 and Th17 responses, plus regulates Treg suppression activity. Our preliminary data shows that TIGIT expressing T cells in mouse kidney are highly activated and produce proinflammatory cytokines after IR. Importantly, mice lacking TIGIT (TIGIT KO) were protected from AKI in IR and Cisplatin AKI models, suggesting detrimental role for TIGIT during AKI. Therefore, understanding TIGIT-mediated inflammatory response during AKI is critical for developing novel AKI therapy and to mitigate kidney adverse effects of immune checkpoint therapies. The central hypothesis of this proposal is that TIGIT promotes proinflammatory functions of kidney T cells and impairs Treg suppression function. To test this hypothesis, we will (Aim 1) investigate phenotypic, functional and transcriptional effects of TIGIT in mouse kidney T cells using in vitro and in vivo approaches. We will further investigate the functional relationship between TIGIT and its co-signaling partners (CD226, CD155) and other co-inhibitory molecules (PD1, CTLA4) in regulating kidney T cell functions at baseline and during AKI. Additionally (Aim 2), we will test the hypothesis that T cell specific TIGIT activity is the major mechanism that drives AKI and impairs repair process using adoptive transfer approaches, in vivo anti-TIGIT agonist/antagonist antibody effects on AKI outcome in WT and TIGIT KO mice and blocking TIGIT signaling in repair phase after established AKI. Finally (Aim 3), we will investigate functional effects of TIGIT expression in human kidney T cells in patients with renal cell carcinoma and live donor biopsies. We will also evaluate TIGIT expression on T cells isolated from ischemic deceased donor kidney samples and transcriptional effects at single cell level in T cells from live donor kidney. Results from these studies will be the first to provide important information on TIGIT mediated effect in kidney T cell functions, therapeutic potential of targeting TIGIT for AKI treatment and set the stage for future pre-clinical and clinical studies.