铁死亡相关特发性肺纤维化特征基因的筛选及功能研究

Identification and functional study of ferroptosis-related genes in idiopathic pulmonary fibrosis

目的:筛选铁死亡(ferroptosis,Fer)相关的特发性肺纤维化(idiopathic pulmonary fibrosis,IPF)特征基因及探讨IPF潜在发病机制。方法:利用WGCNA和ssGSEA构建铁死亡相关的IPF风险模型(Fer-Score),分析不同风险亚型免疫细胞浸润和通路富集。进一步利用LASSO算法对IPF特征的铁死亡相关基因进行筛选,并进行免疫细胞浸润丰度、关键通路评分与特征基因相关性分析。最后通过IPF细胞模型验证特征基因的表达及细胞功能。结果:筛选出9个铁死亡相关IPF基因用于构建Fer-Score。不同Fer-Score亚型的风险评分结果差异具有统计学意义,发现集及验证集的ROC曲线下面积分别为0.9177和0.9375。免疫细胞浸润分析结果显示记忆B细胞、树突状细胞、静息自然杀伤(natural killing,NK)细胞、单核细胞、中性粒细胞与富集分数显著相关。GSEA通路富集分析发现共有24条通路显著富集,其中有7条通路与免疫细胞浸润显著相关。结合LASSO算法和Pearson相关性分析发现MEG8、ALOX15与ADAM23是导致IPF发生的特征基因。细胞实验结果发现ADAM23在IPF细胞模型中显著高表达,敲低ADAM23可抑制脂多糖(lipopolysaccharide,LPS)诱导的BEAS-2B细胞的增殖和迁移。结论:敲低ADAM23可抑制特发性肺纤维化进程,其可能通过调控代谢相关通路影响免疫细胞浸润水平从而导致IPF的发生。

Objective:This study aims to explore the characteristic genes and potential mechanisms of ferroptosis-related idiopathic pulmonary fibrosis(IPF).Methods:To construct a ferroptosis-related IPF risk model(Fer-Score)by using WGCNA and ssGSEA.Subsequently analyzed immune cell infiltration and pathway enrichment in different risk subtypes.The LASSO algorithm was employed to screen ferroptosis-related genes characteristic of IPF and assess the correlation between immune cell infiltration abundance,key pathway scores and characteristic genes.Finally,we validated the expression of characteristic genes and conducted functional studies using an IPF cell model.Results:Nine ferroptosis-related IPF genes were identified for constructing the Fer-Score.The risk scores of different subtypes of Fer-Score showed significant difference,with the model achieving an area under the ROC curve of 0.9177 and 0.9375 for the discovery and validation sets,respectively.Immune infiltration analysis revealed significant correlations between enrichment scores and memory B cells,dendritic cells,resting NK cells,monocytes,and neutrophils.Pathway enrichment analysis identified 24 pathways significantly enriched by GSEA,seven of which were notably related to immune cell infiltration.Combined LASSO algorithm and Pearson correlation analysis identified MEG8,ALOX15,and ADAM23 as characteristic genes contributing to IPF.Cell experiments showed that ADAM23 was significantly overexpressed in the IPF cell model,and knockdown of ADAM23 inhibited LPS induced proliferation and migration of BEAS-2Bs cells.Conclusion:Knockdown of ADAM23 can inhibit the progression of IPF potentially by regulating metabolism-related pathways to influence the infiltration patterns of five types of immune cells.Thereby ADAM23 may contribute to the occurrence of IPF.