Lung Injury Repair by Regulatory T cell LGP2

调节性 T 细胞 LGP2 修复肺损伤

基本信息

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
项目类别：
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

项目摘要

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解决的机制严重的急性肺部炎症。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遇险综合征（ARDS），这是一种毁灭性的疾病，在美国引起严重的病态和死亡率尽管对损伤和炎症进行了调查，但驱动驱动力ARD的炎症，但没有靶向疗法促进它的分辨率。系统激活 - 在肺损伤的小鼠模型中溶解炎症。肺损伤后的Treg功能仍然未知。编码免疫调节蛋白LGP2，尽可能高达新的部位，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基因座的再生。因此，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 pro-Repair功能，并促进急性肺炎症的分辨率。具体目标是支撑：1。定义DNMT-UHRF1复合物在促进LGP2甲基化中的作用 ALI分辨率的TREG；确定LGP2在调节ALI期间的Treg Repair函数中的作用解决方案和3 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。我们还设计了一种RNA干扰策略用突变的DNA质粒增强DHX58的表达。已建立的急性肺损伤的小鼠模型给药），DNA甲基化测序技术和多色流式细胞计这些目标将发现控制急性肺肺肺Treg功能的机制翻译为ARDS的治疗益处。