Identification of Small Airway Epithelium-Related Hub Genes in Chronic Obstructive Pulmonary Disease

Lanlan Lin1,2 *, Guofu Lin1,2 *, Xiaohui Chen,1,2 Hai Lin,1,2 Qinhui Lin,1,2 Yiming Zeng,1– 3 Yuan Xu1– 3 1Department of Pulmonary and Critical Care Medicine, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, People’s Republic of China; 2Respiratory Medicine Center of Fujian Province, Quanzhou, People’s Republic of China; 3Clinical Research Center, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yuan Xu; Yiming Zeng, Department of Pulmonary and Critical Care Medicine, Clinical Research Center, The Second Affiliated Hospital of Fujian Medical University; Respiratory Medicine Center of Fujian Province, Quanzhou, People’s Republic of China, Email yuan_xu0123@fjmu.edu.cn; zeng_yiming@fjmu.edu.cnBackground: Pulmonary small airway epithelia are the primary site of cellular and histological alterations in chronic obstructive pulmonary disease (COPD), while the potential therapeutic hub genes of pulmonary epithelia are rarely identified to elucidate profound alterations in the progression of the disease.Methods: Microarray dataset of GSE11906 containing small airway epithelia from 34 healthy non-smokers and 33 COPD patients was applied to screen differentially expressed genes (DEGs). Weighted gene correlation network analysis (WGCNA) was further used to identify the hub genes related to clinical features. Moreover, single-cell RNA sequencing data from GSE173896 and GSE167295 dataset were applied to explore the expression and distribution of the hub genes. The expression levels of hub genes in epithelial cells stimulated by cigarette smoke extract (CSE) were detected by RT-qPCR.Results: Ninety-eight DEGs correlated with clinical features of COPD were identified via limma and WGCNA. Eight hub genes (including AKR1C3, ALDH3A1, AKR1C1, CYP1A1, GPX2, CBR3, AKR1B1 and GSR) that might exert an antioxidant role in COPD process were identified. Single-cell transcriptomic analysis indicated that the expressions of AKRAC3, ALDH3A1, GPX2, CBR3 and AKR1B1 were significantly increased in the COPD group when compared with the normal group. Moreover, we found that the expression of ALDH3A1 was the most abundantly expressed in ciliated cells. RT-qPCR results indicated that the majority of candidate novel genes were significantly elevated when the epithelial cells were exposed to CSE.Conclusion: Through integrating limma, WGCNA, and protein-protein interaction (PPI) analysis, a total of eight candidate hub genes of pulmonary airway epithelia were identified in COPD. Moreover, single-cell transcriptomic analysis indicated that ALDH3A1 was enriched in ciliated cells, which may provide a new insight into the pathogenesis and treatment of COPD.Keywords: COPD, pulmonary airway epithelium, hub genes, cigarette smoke