Lung Injury Repair by Regulatory T cell LGP2

项目总结 调节性T细胞LGP2对肺损伤的修复作用 这个R01提案的总体目标是研究引导免疫系统解决 严重的急性肺部炎症。这项提案的团队由来自两个主要国家的专家调查人员组成 大学，设计了一项研究战略，将验证新的治疗目标急性呼吸窘迫 综合征(ARDS)，这是一种破坏性的疾病，在美国导致显著的发病率和死亡率。 尽管对导致ARDS的损伤和炎症进行了彻底的调查，但没有针对性的治疗方法促进 它的决心。调节性T细胞(Tregs)--抑制旺盛免疫的CD4+淋巴细胞亚群 系统激活-在小鼠肺损伤模型中化解炎症。然而，促进 肺损伤后的Treg功能仍不清楚。在Tregs中发现的Lgp2基因座的初步数据， 编码免疫调节蛋白LGP2，作为一个新的位点，增强Treg对炎症的反应。 这个位点的DNA甲基化是动态的，并有助于肺损伤后Dhx58基因的抑制。 因此，Lgp2基因座上Treg DNA低甲基化的假说会增加LGP2蛋白水平，增强 Treg具有促修复功能，促进急性肺部炎症的消退。为了验证这一假设，请看以下内容 具体目标是：1.确定Dnmt-uhrf1复合体在促进Lgp2甲基化中的作用 Tregs在ALI中的作用；2.确定LGP2在ALI中调节Treg预修复功能中的作用 3.评价JHU848在促进Treg介导的ALI溶解中的作用。专门测试 我们的假设是，我们将使用转基因小鼠，包括Dhx58缺乏的菌株以及Treg特异性的 Uhrf1缺乏。我们还设计了一种RNA干扰策略来敏锐地敲除培养的 用突变的DNA质粒Tregs和Boost Dhx58表达。这项建议的主要方法包括 建立小鼠急性肺损伤(气管内脂多糖和铜绿假单胞菌)模型 管理)、DNA甲基化测序技术和多色流式细胞术。成就 这些目标将揭示在急性肺损伤缓解过程中控制Treg功能的机制，这可能是 翻译为ARDS的治疗益处。

PROJECT SUMMARY Lung Injury Repair by Regulatory T cell LGP2 The overall objective of this R01 proposal is to investigate mechanisms that direct the immune system to resolve severe acute lung inflammation. The team for this proposal, comprising expert investigators from two leading universities, has designed a research strategy that will validate new therapeutic targets acute respiratory distress syndrome (ARDS), which is a devastating condition that causes significant morbidity and mortality in the U.S. Despite thorough investigation into the injury and inflammation that drive ARDS, no targeted therapies promote its resolution. Regulatory T cells (Tregs) — a subset of CD4+ lymphocytes that suppress exuberant immune system activation—resolve inflammation in mouse models of lung injury. However, the mechanisms that promote Treg function following lung injury remain unknown. Preliminary data identified in Tregs the Lgp2 locus, which encodes the immune regulatory protein LGP2, as a novel site that augments Treg responses to inflammation. DNA methylation at this site is dynamic and contributes to repression of the Dhx58 locus following lung injury. Thus, the hypothesis of Treg DNA hypomethylation at the Lgp2 locus will increase LGP2 protein levels, enhance Treg pro-repair function, and promote resolution of acute lung inflammation. To test this hypothesis the following Specific Aims are proposed: 1. Define the role of the Dnmt-Uhrf1 complex in promoting methylation of Lgp2 in Tregs during ALI resolution; 2. Determine the role of LGP2 in regulating Treg pro-repair function during ALI resolution; and 3. Evaluate the role of JHU848 in promoting Treg-mediated ALI resolution. To specifically test our hypothesis we will employ transgenic mice including strains with Dhx58 deficiency as well as Treg-specific Uhrf1 deficiency. We have also designed an RNA interference strategy to acutely knock down Dhx58 in cultured Tregs and boost Dhx58 expression with mutant DNA plasmids. Major methods for this proposal include established mouse models of acute lung injury (intratracheal lipopolysaccharide and Pseudomonas aeruginosa administration), DNA methylation sequencing techniques, and multicolor flow cytometry. Accomplishment of these aims will uncover mechanisms controlling Treg function during resolution of acute lung injury that could be translated for therapeutic benefit in ARDS.