Time-series transcriptomic analysis of lung fibroblasts in bleomycin- and silica-induced pulmonary fibrosis

Pulmonary fibrosis (PF) is an intractable disorder with a poor prognosis. Although lung fibroblasts play central roles in PF, their key regulatory molecules remain unclear. To identify PF pathology-associated gene modules and their hub TFs in activated lung fibroblasts, we performed time-course transcriptome analysis of the fibroblasts purified from the lungs of bleomycin- and silica-treated Col1a2-GFP reporter mice. To clarify transcriptomic landscape of lung fibroblasts throughout PF initiation, progression, and resolution, we induced reversible or progressive PF in Col1a2-GFP transgenic mice by bleomycin or silica aspiration, respectively. We purified lineage (CD45, CD31, EpCAM, CD146, and Ter119)– Col-GFP+ lung fibroblasts by flow-cytometry at 0, 7, 14, and 63 days post-induction in both models and performed transcriptome analysis by 3′SAGE-seq using Ion Proton sequencer.

肺纤维化（Pulmonary fibrosis, PF）是一类预后不良的难治性疾病。尽管肺成纤维细胞在PF的病理进程中发挥核心作用，但其关键调控分子仍未明确。为鉴定活化肺成纤维细胞中与PF病理相关的基因模块及其枢纽转录因子（transcription factor, TF），我们对经博莱霉素与二氧化硅处理的Col1a2-GFP报告基因小鼠肺部纯化分离的成纤维细胞开展了时序转录组分析。为阐明肺纤维化从起始、进展至消退全程中肺成纤维细胞的转录组全景，我们分别通过博莱霉素吸入给药与二氧化硅吸入给药，在Col1a2-GFP转基因小鼠中构建了可逆性与进展性PF模型。在两种模型中，我们分别于诱导后第0、7、14及63天，通过流式细胞术分选获得谱系标记阴性（CD45、CD31、EpCAM、CD146及Ter119）的Col-GFP阳性肺成纤维细胞，并采用Ion Proton测序仪通过3′SAGE-seq（3'端基因表达连续分析测序）完成转录组测序分析。