LncRNA regulates lung inflammation

摘要 肺部感染是全球公共卫生的主要负担，也是全球人口死亡的主要原因。 美国。由革兰氏阴性(G-)细菌引起的感染具有特别令人关注的特征， 比如在获得抗生素耐药性方面非常有效。肺泡巨噬细胞(AM)形成第一条线 通过保护性炎症反应在肺部防御微生物病原体。高死亡率和 细菌感染后的发病率通常是由于杀菌剂和细菌之间的宿主防御失衡造成的 过度的炎症反应，导致组织损伤。尽管进行了多年的研究，入门仪式， G-细菌感染残留时炎性肺反应的传播和调节 没有完全探索过。直到最近人们才认识到，长的非编码RNA(LncRNA)是 在包括巨噬细胞在内的各种免疫细胞中广泛表达，但人们对此知之甚少 他们的职能角色。我们筛选了气管内注入脂多糖后小鼠肺组织中lncRNA的表达， 肺炎克雷伯菌(肺炎克雷伯菌)和大肠埃希氏菌(E.coli.)LncRNA连锁1显示出最大的 在处理组中诱导表达。在功能上，lincenc1促进经典的激活(M1) 巨噬细胞和促炎细胞因子的分泌。使用特定的删除AMS中的链接1 体内反义寡核苷酸(ASO)可显著减少内毒素诱导的白细胞浸润和肺泡形成 水肿，与对照组比较。此外，使用ASO抑制lincenc1下调 暴露后肺泡灌洗液中多种炎性细胞因子和趋化因子(IL-1β、IL-6、CxCl1和CxCl2)的检测 致内毒素。从机制上讲，我们发现在内毒素存在的情况下，lincenc1与NLRP3组分共定位， 提示Lincenc1对NLRP3炎症因子的激活具有调节作用。基于我们发布的和 初步数据，我们的中心假设是lncRNA lincenc1促进G-细菌/内毒素诱导的肺 炎症通过NLRP3炎性小体组装促进巨噬细胞的经典激活和 激活。我们提出了三个具体目标：具体目标一：确定lncRNA连锁1在 巨噬细胞的体外激活。特定目的II：确定lincenc1在体内肺部炎症中的作用。 具体目的III：确定lincenc1是否在体内作为潜在的治疗靶点。

ABSTRACT Lung infections place a major burden on public health worldwide and are the leading cause of death in the United States. Infections caused by gram-negative (G-) bacteria have features that are of particular concern, such as being highly efficient at acquiring antibiotic resistance. Alveolar macrophages (AMs) form the first line of defense in lungs toward microbial pathogens via protective inflammatory responses. The high mortality and morbidity after bacterial infection often result from an imbalance in host defense between bactericidal and an excessive inflammatory response that leads to tissue damage. Despite years of research, the initiation, propagation and regulation of inflammatory lung responses in the presence of G- bacterial infection remains incompletely explored. Only recently has it been recognized that long non-coding RNAs (lncRNA) are extensively expressed in various immune cells including the macrophages, but very little has been known about their functional roles. We screened lncRNA expressions in mouse lungs after intra-tracheal instillation of LPS, Klebsiella pneumoniae (K.pneumoniae) and Escherichia coli (E.coli). LncRNA lincenc1 exhibited the greatest induction in expression among treated groups. Functionally, lincenc1 promotes the classical activation (M1) of macrophages and the secretion of pro-inflammatory cytokines. Deletion of lincenc1 in AMs using the specific antisense oligonucleotides (ASO) in vivo significantly reduces the LPS-induced leukocyte infiltration and alveolar edema, when compared with the control group. Furthermore, inhibition of lincenc1 using ASO down-regulates multiple inflammatory cytokines and chemokines (Il-1β, Il-6, Cxcl1 and Cxcl2), detected in BALF after exposure to LPS. Mechanistically, we found that lincenc1 co-localizes with NLRP3 components in the presence of LPS, suggesting a regulatory role of lincenc1 on NLRP3 inflammasone activation. Based on our published and preliminary data, our central hypothesis is that lncRNA lincenc1 facilitates G-bacteria/LPS induced lung inflammation via promoting classical activation of macrophages through NLRP3 inflammasome assembly and activation. We propose three specific aims: Specific Aim I: To determine the role of lncRNA lincenc1 in macrophage activation in vitro. Specific Aim II: To determine the role of lincenc1 in lung inflammation in vivo. Specific Aim III: To determine whether lincenc1 serves as a potential therapeutic target in vivo.