Lung Injury Repair by Regulatory T cell LGP2

调节性 T 细胞 LGP2 修复肺损伤

• 批准号: 9309225
• 负责人: Franco R D'Alessio
• 金额: $ 40.89万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9309225
• 关键词: Acute; Acute Lung Injury; Adoptive Transfer; Adult Respiratory Distress Syndrome; Alveolar; Attenuated; Biological Availability; CD4 Positive T Lymphocytes; Cell Therapy; Cells; Complex; Cytosine; DNA; DNA Methylation; DNA Modification Methylases; Data; Deoxycytidine; Disease; Dose; Drug Delivery Systems; Drug Targeting; Epigenetic Process; Epithelial; FOXP3 gene; Flow Cytometry; Gene Expression Profile; Genes; Genetic Transcription; Genomics; Glucocorticoids; Goals; Guanine; IL2RA gene; Immune; Immune system; In Vitro; Inflammation; Inflammatory Response; Influenza; Injury; Investigation; Lipopolysaccharides; Lung; Lung Inflammation; Lymphoid Cell; Malignant Neoplasms; Mediating; Methods; Methylation; Modeling; Morbidity - disease rate; Mus; Outcome; Pathway interactions; Pattern; Phase; Phenotype; Physiological; Pre-Clinical Model; Prodrugs; Proteins; Pseudomonas aeruginosa; RNA Interference; Regulation; Regulatory T-Lymphocyte; Repression; Research; Research Personnel; Resolution; Role; Site; Site-Directed Mutagenesis; Streptococcus pneumoniae; Syndrome; Techniques; Testing; Therapeutic; Therapeutic Index; Toxic effect; Transgenic Mice; Translating; Treatment-related toxicity; Universities; Viral; Virus Diseases; Work; clinical efficacy; design; experience; functional status; genetic regulatory protein; genome-wide; healing; improved; improved outcome; in vitro testing; in vivo; inflammatory lung disease; injured; injury and repair; insight; knock-down; lung injury; lung repair; macrophage; methylation pattern; mortality; mouse model; mutant; new therapeutic target; novel; novel therapeutics; outcome forecast; overexpression; plasmid DNA; repaired; response; targeted treatment; therapy design; transcription factor; transcriptome sequencing

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.

项目摘要 调节T细胞LGP2修复肺损伤 该R01提案的总体目标是调查指导免疫系统解决的机制 严重的急性肺部感染。该提案的团队，完成了来自两个领先的专家调查员 大学设计了一种研究策略，该策略将验证新的治疗目标急性呼吸障碍 综合征（ARDS），这是一种毁灭性的疾病，在美国引起明显的发病率和死亡率 尽管对驱动ARDS的伤害和感染进行了彻底的调查，但尚无靶向疗法促进 它的解决方案。调节性T细胞（Tregs） - CD4+淋巴细胞的子集，可抑制激增免疫 系统激活 - 在肺损伤的小鼠模型中溶解注射。但是，促进的机制 肺损伤后的Treg功能仍然未知。 Tregs在LGP2基因座中确定的初步数据 编码免疫调节蛋白LGP2，是一个新的部位，增强了Treg对炎症的反应。 该部位的DNA甲基化是动态的，并有助于肺损伤后DHX58基因座的表达。 这，在LGP2基因座上Treg DNA降压甲基化的假设将增加LGP2蛋白水平，增强 Treg促进功能，并促进急性肺注射的分辨率。为了检验这一假设以下 提出了具体目的：1。定义DNMT-UHRF1复合物在促进LGP2甲基化中的作用 阿里解决期间的特雷格； 2。确定LGP2在调节ALI期间调节Treg Pro-Repair函数中的作用 解决;和3。评估JHU848在促进Treg介导的ALI分辨率中的作用。专门测试 我们的假设我们将采用转基因小鼠，包括DHX58缺乏症以及特雷格特异性的菌​​株 UHRF1缺乏症。我们还设计了一种RNA干扰策略，以急性地击倒培养的DHX58 用突变的DNA质粒增强DHX58的表达。该建议的主要方法包括 已建立的急性肺损伤的小鼠模型（气管内脂多糖和铜绿假单胞菌 给药），DNA甲基化测序技术和多色流式细胞仪。成就 这些目标将发现控制Treg功能的机制，这可能是 翻译为ARDS的治疗益处。