Single-Cell Transcriptomic Profiling Reveals Molecular Alterations in the Airway Epithelial Cells of Children with Wheezing Infected by Respiratory Syncytial Virus

Abstract

This study aimed to explore the molecular alterations in airway epithelial cells in children with wheezing infected by respiratory syncytial virus (WheezeRSV). This study extracted the single-cell sequencing data of two control and wheezeRSV samples from GSE286262 dataset. The monocle2 was performed for analyzing evolution process of basal cells along disease progression. Kyoto encyclopedia of genes and genomes and gene ontology were carried out to identify the function enrichment of differentially expressed genes (DEGs) between WheezeRSV and control groups. Additionally, human nasal epithelial cells (HNECs) were harvested, and quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect the expression levels of key genes. Six cell types including ciliated cells, KRT5 + basal cells, TP63 + basal cells, KRT80 + epithelial cells, goblet cells, and club cells. The differentiation trajectory analysis on basal cells indicated two distinct branches including ciliary assembly and keratinization. Then, function analysis indicated that ciliated cells were involved in leukocyte transendothelial migration, cellular senescence and angiogenesis. Goblet cells were associated with IL-17 signaling pathway, apoptosis, and ferroptosis, and club cells were enriched in the apoptotic process and innate immune response. The qRT-PCR results revealed that the mRNA expression levels of TEKT2, SPRR3, ITGB1, CDKN1A, ACSL1, and CXCL8 were markedly upregulated in the WheezeRSV group (p < 0.01). Basal cells differentiated into keratinocytes and ciliated cells as the disease progresses, thereby enhancing defense and repairing the epithelial barrier. Meanwhile, wheezeRSV enhanced the activity of apoptosis and inflammatory response of ciliated cells, goblet cells, and club cells.