Resident Memory T cells in Chronic Kidney Disease

项目摘要 慢性肾脏疾病（CKD）是一种具有显著发病率和死亡率的疾病，其影响数百万人， 美国人并没有表现出改善的迹象。因为没有临床批准的治疗方法可以 阻止或逆转CKD向终末期肾病的进展，需要研究来阐明新的 分子和细胞靶点。这个项目是基于科学的前提，炎症和纤维化是 CKD进展的关键因素。虽然最近的研究强调了免疫系统的重要性， 尽管在CKD的免疫系统中，驻留记忆T细胞（TRM）的具体贡献仍然未知。我们有 建立并验证了马兜铃酸（AA）诱导的模拟临床CKD的CKD小鼠模型。 连续五天的AA注射导致肾功能持续下降，表现为肾功能下降。 6周后肾小球滤过率和血清肌酐升高。肾脏的流式细胞术显示， 在第一次AA注射的两周内主要是淋巴细胞浸润，CD 4和CD 8 T细胞是主要的淋巴细胞浸润。 最丰富的。这些T细胞持续至少6周，与肾功能的持续下降相一致。 此外，这些小鼠肾脏中的大量T细胞表达促炎和纤维化蛋白。 细胞因子，以及CD 103，TRM的标志性上皮粘附分子。TRM被描述为 在其他慢性疾病中，它们是不必要的炎症的贡献者， 在人类肾脏中。因此，我们的中心假设是，TRM介导的进展，慢性肾脏病通过 炎症反应，促进肾损伤和纤维化，同时抑制修复。本提案的目的是 将1）确定TRM的激活状态和细胞因子谱以及它们对肾脏的贡献， 炎症和纤维化;和2）确定CD 103对T细胞-上皮粘附的必要性及其对 T细胞定位于受损肾脏，以及其对TRM促炎状态的必要性。实现 这些目标，我们将利用基因敲除模型，过继免疫细胞转移，流式细胞术， 免疫荧光显微镜和药物干预在体外和体内。成功 建议的研究的执行将阐明CKD的基本机制，并可提供 CKD治疗新靶点的临床证据。这符合NIDDK的使命，因为 该提案的重点是阐明进行性肾病的治疗方法。的实验室 赞助商沿着他们在肾脏学和免疫学方面的专业知识，将提供理想的培训环境， 开展拟议的研究并推进PI的职业生涯。接受的培训将增进知识， 批判性思维，技术和专业技能，将需要PI的过渡到独立的 学术研究和建立他在心肾疾病领域的利基。

Project Summary Chronic kidney disease (CKD) is a disorder with significant morbidity and mortality which affects millions of Americans and shows no sign of improvement. Because there are no clinically approved therapeutics that can halt or reverse the progression of CKD to end stage kidney disease, research is needed to elucidate novel molecular and cellular targets. This project is based on the scientific premise that inflammation and fibrosis are key components in the progression of CKD. While recent research has highlighted the importance of the immune system in CKD, the specific contributions of resident memory T cells (TRMs) remain unknown. We have established and validated an aristolochic acid (AA)-induced mouse model of CKD which mimics clinical CKD. Five consecutive days of AA injections lead to a sustained decline in kidney function, indicated by reduced glomerular filtration rate and increased serum creatinine six weeks later. Flow cytometry of kidneys reveals a predominantly lymphocytic infiltrate within two weeks of the first AA injection, CD4 and CD8 T cells being the most abundant. These T cells persist for at least 6 weeks, coinciding with the persistent decline in kidney function. Moreover, a large population of T cells in the kidneys of these mice express pro-inflammatory and fibrotic cytokines, as well as CD103, a hallmark epithelial adhesion molecule of TRMs. TRMs have been described as contributors to unwanted inflammation in other chronic diseases and their presence has been descriptively noted in human kidneys. Thus, our central hypothesis is that TRMs mediate the progression of CKD through an inflammatory response that promotes kidney injury and fibrosis while inhibiting repair. The aims of this proposal will 1) determine the activation status and cytokine profiles of TRMs as well as their contributions to kidney inflammation and fibrosis; and 2) determine CD103’s necessity for T cell-epithelial adhesion and its regulation of T cell localization to injured kidneys, as well as its necessity for the TRM pro-inflammatory state. To achieve these goals, we will utilize genetic knockout models, adoptive immune cell transfer, flow cytometry, immunofluorescence microscopy, and pharmaceutical interventions both in vitro and in vivo. Successful execution of the proposed studies will illuminate basic mechanisms involved in CKD and could provide pre- clinical evidence for novel targets in the treatment of CKD. This is in line with the mission of the NIDDK, as this proposal focuses on the elucidation of treatments for progressive kidney disease. The laboratories of the sponsors along with their expertise in nephrology and immunology will provide an ideal training environment to carry out the proposed studies and advance the PI’s career. The training received will advance the knowledge, critical thinking, technical, and professional skills that will be required for the PI’s transition into independent academic research and establishment of his niche in the field of cardiorenal disease.