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 调节性T细胞LGP 2对肺损伤的修复 这项R 01提案的总体目标是研究指导免疫系统解决 严重的急性肺部炎症。该提案的小组由来自两个主要国家的专家调查员组成， 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 急性呼吸窘迫综合征（ARDS），这是一种在美国造成严重发病率和死亡率的破坏性疾病。 尽管对驱动ARDS的损伤和炎症进行了彻底的调查，但没有靶向治疗促进 其决议。调节性T细胞（Treg）-抑制旺盛免疫反应的CD 4+淋巴细胞亚群 系统激活-消除小鼠肺损伤模型中炎症。然而，促进 肺损伤后Treg的功能仍不清楚。初步数据表明，在Tumor中发现了Lgp 2基因座， 编码免疫调节蛋白LGP 2，作为增强Treg对炎症反应的新位点。 该位点的DNA甲基化是动态的，并有助于肺损伤后Dhx 58位点的抑制。 因此，在Lgp 2基因座处Treg DNA低甲基化的假设将增加LGP 2蛋白水平，增强LGP 2基因的表达。 Treg具有促修复功能，促进急性肺部炎症消退。为了验证这一假设， 具体目标是：1。确定Dnmt-Uhrf 1复合物在促进Lgp 2甲基化中的作用， ALI消退期间的TBI; 2.确定LGP 2在ALI期间调节Treg促修复功能中的作用 决议; 3。评价JHU 848在促进Treg介导的ALI消退中的作用。专门测试 我们的假设是，我们将使用转基因小鼠，包括Dhx 58缺陷型以及Treg特异性 Uhrf 1缺陷。我们还设计了一种RNA干扰策略，在培养的细胞中急性敲低Dhx 58， 用突变体DNA质粒转染和加强Dhx 58表达。本提案的主要方法包括 建立小鼠急性肺损伤模型（内毒素和铜绿假单胞菌 施用）、DNA甲基化测序技术和流式细胞术。完成 这些目标将揭示在急性肺损伤消退过程中控制Treg功能的机制， 转化为治疗ARDS的益处。