Effect of FN14KO on mouse lung immune cells after bleomycin injury

TWEAK, also known as TNFSF12, is a multifunctional cytokine that controls many cellular activities in tissue repair and disease by binding to its receptor Fn14.TWEAK-Fn14 signaling has been shown to induce canonical and noncanonical NFkB signaling, triggering pro-inflammatory responses and leukocyte infiltration.In the bleomycin-induced lung fibrosis model, Fn14 KO mice were found to induce a more severe fibrotic phenotype. To explore the effect of Fn14 KO on immune cell populations in this model, we performed single-cell sequencing. Overall design: Immune cells from six group of mice(WT saline, FN14KO saline, WT bleomycin D7, FN14KO bleomycin D7, WT bleomycin D14, FN14KO bleomycin D14) were isolated by FACS(CD45+ EPCAM- CD31-)and analyzed using scRNAseq.

TWEAK（TNFSF12）是一种多功能细胞因子，可通过结合其受体Fn14，调控组织修复与疾病进程中的多种细胞活动。已有研究表明，TWEAK-Fn14信号通路可诱导经典与非经典核因子κB（NFκB）信号活化，触发促炎反应及白细胞浸润。在博莱霉素诱导的肺纤维化模型中，研究发现Fn14敲除（knockout, KO）小鼠可表现出更为严重的纤维化表型。为探究该模型中Fn14敲除对免疫细胞群的影响，本研究开展了单细胞RNA测序（single-cell RNA sequencing, scRNAseq）。总体实验设计：通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS，标记CD45+ EPCAM- CD31-）分离6组小鼠的免疫细胞，具体分组为野生型（Wild Type, WT）生理盐水组、FN14KO生理盐水组、野生型（WT）博莱霉素处理7天组、FN14KO博莱霉素处理7天组、野生型（WT）博莱霉素处理14天组、FN14KO博莱霉素处理14天组，随后利用scRNAseq对样本进行分析。