大气细颗粒物致气道重塑和肺气肿的非编码RNA相关分子标志筛选及早期预警体系研究

The identification of differentially expressed non-coding RNAs (ncRNA) plays an important role to establish the early-warning system to fine particulate matter induced airway remodeling and emphysema. Our previous researches have focused on the toxic components of fine particulate matter (PM2.5) and its pulmonary toxicity mechanism. In the current study, we are trying to identify PM2.5 exposure-related differentially expressed ncRNAs and the association between PM2.5 exposure and COPD, based on panel study. The molecular mechanism involved in PM2.5 induced airway remodeling and emphysema, such as EMT and autophagy, will be studied by in vitro and in vivo models, which help to identify the key molecules in these pathways. We use bioinformation technique to construct the ncRNA-key molecules networks, meanwhile the specific AOP involved in PM2.5 exposure induced COPD. Detection of molecular initial events, like differentially expressed ncRNAs, in AOP are essential for the early-warning system, therefore, we are trying to design a paper-based microfluidic analytical device for the high-throughput detection of ncRNA bionarkers in fielding study.Our project would facilitate the integrative understanding of mechanisms involved in PM2.5-induced airway remodeling and emphysema, as well as the establishment of early-warning system, ultimately to prevent against PM2.5 exposure induced COPD.

PM2.5所致气道重塑和肺气肿病理过程中，调控关键分子表达水平的非编码RNA（ncRNA）的识别对建立早期预警体系尤为重要。基于项目组前期对PM2.5毒性组分解析及机制研究，拟通过定组研究，筛选出与PM2.5暴露相关的差异表达ncRNA，同时明确PM2.5暴露与COPD的关联。结合细胞及动物模型探究PM2.5暴露致气道重塑和肺气肿的关键分子机制，包括上皮-间充质转化（EMT）及自噬，发现通路中的关键分子；生物信息学构建ncRNA关键分子调控网络及特异性有害结局通路（AOP）网络，识别起始分子事件（MIE）。将纸微流控芯片技术应用于所识别的MIE即ncRNA分子标志检测，最终实现PM2.5暴露相关的COPD早期预警。本研究不仅将阐明PM2.5暴露致气道重塑和肺气肿的早期分子机制，更将提供ncRNA分子快速、标准、高通量的现场检测手段，为促进PM2.5的早期预防和早期预警提供新方法及新思路。

本项目利用细胞、动物模型以及人群定组研究，识别PM2.5致气道重塑及肺气肿病理过程中的关键非编码RNA分子，构建ncRNA关键分子调控网络及特异性有害结局通路（AOP），全面评价表观遗传修饰作为PM2.5暴露所致的气道重塑和肺气肿效应标志的可行性及早期预警体系，完成如下目标：（1）采用人群定组研究探讨PM2.5暴露与COPD的效应关系。（2）建立体内外染毒模型，阐明PM2.5暴露致气道重塑和肺气肿的关键分子机制和早期非编码RNA效应分子。（3）构建ncRNA分子调控网络及特异性有害结局通路，开发ncRNA分子微流控芯片检测技术，实现PM2.5暴露相关的COPD早期预警。研究结果发现：（1）在人群水平，柴油尾气来源的1-NP内暴露水平显著升高，其代谢产物水平与COPD患者血清IL-6水平正相关，机制发现1-NP通过诱导细胞自噬发挥细胞毒性作用，牛磺酸可以缓解1-NP诱导的肺气肿。（2）PM2.5暴露诱导COPD患者血浆中circ_5045表达增加，circRNA_5045与ELANE、PRDX2结合，促进COPD样损害；PM2.5暴露激活miR-382-5p/CXCL12/MMP9通路促进肺部炎症；PM2.5暴露诱导lncRNA-TUG1/miR-222-3p/CELF1通路激活致气道高反应性。（3）构建PM2.5致COPD特异性AOP网络，阐明非编码RNA表达谱的改变作为分子启动事件，氧化应激和炎症通路激活诱导肺部炎症、上皮细胞-间充质转化致气道重塑、肺气肿为关键事件。（4）构建基于机器学习预测模型，鉴别COPD患者对PM2.5污染响应因素，评估PM2.5污染致健康危害的风险。（5）利用PM2.5诱导的HBE细胞恶性转化模型，关键分子ACLY参与的柠檬酸盐代谢通路激活，ACLY敲减可延缓细胞恶性转化进程。项目负责人受邀参加环境诱变剂国际会议，中华预防医学卫生毒理分会，第五届全国细胞外囊泡大会等本领域内重要学术会议，并做大会及分会场报告。在本项目的资助下项目负责人获得2020年度国家杰出青年基金资助，项目成员获批国自然青年项目2项、国自然面上项目1项，培养博士研究生4名，硕士研究生5名。发表SCI论文13篇、中文2篇。微流控芯片成果已申请国家发明专利2项