Investigating CD4 T regulatory cells during rapid cystogenesis

研究快速囊肿发生过程中的 CD4 T 调节细胞

• 批准号: 9891171
• 负责人: Kurt A Zimmerman
• 金额: $ 15.32万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9891171
• 关键词: Adoptive Transfer; Adult; Affect; Antibodies; B-Lymphocytes; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Carrier Proteins; Cell Count; Cell physiology; Cells; Cilia; Clinic; Cyst; Cystic Kidney Diseases; Cystic kidney; Data; Development; Disease; End stage renal failure; Environmental Risk Factor; Epithelial; Epithelial Cell Proliferation; Epithelium; FOXP3 gene; Flow Cytometry; Foxes; Functional disorder; Future; Genetic; Goals; Growth; Healthcare; Healthcare Industry; Human; Immune; Inflammatory; Injury; Injury to Kidney; Interleukin-10; Intervention; Kidney; Knowledge; Link; Meckel-Gruber syndrome; Mediating; Modeling; Mus; Mutation; PKD2 protein; Patients; Pharmaceutical Preparations; Pharmacology; Population; Process; Proteins; Rag1 Mouse; Regulatory T-Lymphocyte; Renal function; Reperfusion Injury; Reporter; Reporting; Severities; Testing; Time; Wild Type Mouse; care burden; cell motility; cytokine; disease-causing mutation; experience; imaging approach; injured; interleukin-10 receptor; intravital imaging; mouse model; mutant; new therapeutic target; novel; polycystic kidney disease 1 protein; recruit; repaired; targeted treatment; transcription factor

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.

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。这一系列的疾病是由蛋白质突变引起的 纤毛形成（鞭毛内转运蛋白88，Ift 88）或功能（多囊蛋白1，Pkd 1;多囊蛋白2， Pkd2）。此外，患有这些疾病的患者会经历缓慢的局灶性囊肿形成和快速、严重的囊肿形成。 囊肿进展。来自小鼠模型的数据表明纤毛相关蛋白（Ift 88、Pkd 1、Pkd 2）的丢失导致 在缓慢的局灶性囊肿形成中，提示快速囊肿形成需要额外的环境因素， 在人类患者中经常观察到。这一假设得到了多个实验室数据的支持，包括我们自己的实验室， 表明肾损伤促进纤毛功能障碍小鼠的快速膀胱形成。重要的是，肾损伤 促进野生型小鼠中免疫细胞的积累。我们实验室和其他实验室的数据表明， 纤毛功能障碍的小鼠中细胞积累进一步增强和延长，表明持续的 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，对损伤后促进囊性疾病的免疫细胞类型知之甚少， 代表了知识的差距。本申请中概述的初步数据表明，所有基因的遗传缺失都可能导致基因缺失。 适应性免疫细胞（CD 4 T细胞、CD 8 T细胞和B细胞）或仅CD 4 T细胞的药理学耗竭 减少纤毛功能障碍小鼠损伤后的囊肿严重程度，表明CD 4 T细胞可能是关键 肾损伤和快速膀胱形成之间的联系。为了鉴定在这一过程中重要的CD 4 T细胞亚型， 我们在损伤后的不同时间点对对照和纤毛突变肾进行流式细胞术分析。 我们的初步数据表明，纤毛功能障碍的小鼠增加和持续积累的 CD 4 + Foxp 3+调节性T细胞（T细胞）。这些数据导致了总体假设， 损伤后持续的Tcl 4积累导致长期的和异常的上皮细胞增殖 以及纤毛功能障碍小鼠的快速囊肿扩张。此应用程序的目标是识别 快速囊肿形成和调节性T细胞之间的机械联系。我们的假设将被测试与 具体目标如下：1。为了验证纤毛功能障碍导致持续和/或增强的 在肾损伤后邻近扩张的囊肿的区域中TdR的积累。2.为了检验这一假设 在条件性Ift 88小鼠中，TGFAP通过分泌IL-10（一种细胞因子）促进损伤诱导囊肿扩张 它主要由Teptide产生，以前曾报道过驱动囊性上皮细胞 增殖从该提案中收集的数据将用于开发新的免疫细胞调节药物， 影响快速进展性囊性肾患者的Treg数量和功能（产生IL-10的能力） 疾病