Investigating CD4 T regulatory cells during rapid cystogenesis

项目摘要 囊性肾病影响1：500人，占所有终末期肾病患者的5%-10% 这是一个巨大的医疗负担。这一系列疾病是由蛋白质突变引起的。 纤毛形成(鞭毛内转运蛋白88，Ift88)或功能所必需的(Polycystin 1，PKD1；Polycsytin 2， PKD2)。此外，患有这些疾病的患者还会经历一段时间的慢性局灶性囊肿期和快速、重度囊肿期。 囊肿性进展。来自小鼠模型的数据表明纤毛相关蛋白(Ift88，PKD1，PKD2)的丢失导致 慢性局灶性囊变提示快速囊变需要额外的环境因素 通常在人类病人身上观察到。这一假设得到了来自多个实验室的数据的支持，包括我们自己的 表明肾脏损伤促进了纤毛功能障碍小鼠的快速囊变。重要的是，肾脏损伤 促进免疫细胞在野生型小鼠体内的积累。来自我们实验室和其他实验室的数据表明，免疫 纤毛功能障碍小鼠细胞堆积进一步增强和延长，提示持续性 免疫细胞积聚可能是纤毛功能障碍小鼠损伤后迅速形成囊肿的原因。 然而，损伤后促进囊性疾病的免疫细胞类型还知之甚少。 代表着知识上的差距。本申请中概述初步数据显示所有基因缺失 适应性免疫细胞(CD4T细胞、CD8T细胞和B细胞)或仅CD4T细胞的药物耗竭 降低纤毛功能障碍小鼠损伤后的囊肿度提示CD4T细胞可能是关键 肾脏损伤和快速囊变之间的联系。为了确定在这一过程中重要的CD4T细胞亚型， 我们在损伤后的不同时间点对对照肾脏和纤毛突变肾脏进行了流式细胞术分析。 我们的初步数据表明，纤毛功能障碍的小鼠增加并持续积累 CD4+Foxp3+T调节细胞(Tregs)。这些数据导致了总体假设的增加和 损伤后Tregs的持续积聚会导致长时间的异常上皮增殖 纤毛功能障碍小鼠的囊泡迅速扩张。此应用程序的目标是标识 快速成囊和T调节细胞之间的机制联系。我们的假设将通过 具体目标如下：1.检验纤毛功能障碍导致持续性和/或增强性的假设 肾损伤后Tregs积聚在膨胀性囊肿附近。2.检验假设 Tregs通过分泌细胞因子IL-10促进条件性Ift88小鼠损伤诱导的囊性扩张 它主要是由Tregs产生的，之前有报道称它可以驱动囊性上皮细胞 扩散。从这项提案中收集的数据将用于开发新的免疫细胞调节药物， 快速进展性囊性肾炎对Treg细胞数量和功能(产生IL-10能力)的影响 疾病。

Project Abstract Cystic kidney diseases affect 1:500 people and account for 5-10% of all patients with end stage renal disease representing a significant health care burden. This spectrum of disorders is caused by mutations in proteins required for cilia formation (Intraflagellar transport protein 88, Ift88) or function (Polycystin 1, Pkd1; Polycsytin 2, Pkd2). In addition, patients with these disorders experience periods of slow focal cyst formation and rapid, severe cyst progression. Data from mouse models indicate that loss of cilia related proteins (Ift88, Pkd1, Pkd2) results in slow focal cyst formation suggesting that additional environmental factors are required for rapid cystogenesis often observed in human patients. This hypothesis is supported by data from multiple labs, including our own, indicating that renal injury promotes rapid cystogenesis in mice with cilia dysfunction. Importantly, renal injury promotes the accumulation of immune cells in wild type mice. Data from our lab and others indicate that immune cell accumulation is further enhanced and prolonged in mice with cilia dysfunction suggesting that persistent immune cell accumulation may be the cause of rapid cyst formation following injury in mice with cilia dysfunction. However, the type of immune cell that promotes cystic disease following injury is poorly understood and represents a gap in knowledge. The preliminary data outlined in this application show that genetic deletion of all adaptive immune cells (CD4 T cells, CD8 T cells, and B cells) or pharmacological depletion of just CD4 T cells reduces cyst severity following injury in mice with cilia dysfunction suggesting that CD4 T cells may be the critical link between renal injury and rapid cystogenesis. To identify the subtype of CD4 T cell important in this process, we performed flow cytometry analysis of control and cilia mutant kidneys at various time points following injury. Our preliminary data indicate that mice with cilia dysfunction have increased and persistent accumulation of CD4+ Foxp3+ T regulatory cells (Tregs). These data led to the overall hypothesis that increased and persistent accumulation of Tregs following injury causes prolonged and abnormal epithelial proliferation and rapid cyst expansion in mice with cilia dysfunction. The goal of this application is to identify the mechanistic link between rapid cystogenesis and T regulatory cells. Our hypotheses will be tested with the following specific aims: 1. To test the hypothesis that cilia dysfunction causes persistent and/or enhanced accumulation of Tregs in regions adjacent to expanding cysts following renal injury. 2. To test the hypothesis that Tregs promote injury induced cyst expansion in conditional Ift88 mice through secretion of IL-10, a cytokine that is predominantly produced by Tregs and has previously been reported to drive cystic epithelial cell proliferation. Data collected from this proposal will be used to develop novel immune cell modulating drugs that affect Treg number and function (ability to produce IL-10) in patients with rapidly progressive cystic kidney disease.