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Regulatory Effects of Smoking Cessation on the Cellular Microenvironment and Differentially Expressed Genes in Precancerous Lesions of Pulmonary Nodules in Mice based on Single-Cell RNA-Sequencing and Immune Repertoire-Sequencing

ABSTRACT: Smoking cessation is known to reduce lung cancer progression, but its impact on precancerous lesions remains largely unexplored. In this study, a smoking-induced mouse model of pulmonary precancerous lesions was established using 6-week benzo[a]pyrene (B[a]P) exposure. Lung nodules were identified through micro-MRI imaging and histopathological evaluation by H&E staining and fluorescence microscopy. Smoking cessation interventions were applied for 4 and 8 weeks following lesion confirmation. Nodule size was quantified with Generic Medical Imaging software. Single-cell RNA sequencing (scRNA-seq) and immune repertoire sequencing (IR-seq) were performed to analyze immune cell composition, intercellular communication pathways, and receptor diversity. These approaches provide a comprehensive framework for investigating the cellular and molecular changes associated with smoking cessation in early lung lesion development.

ORGANISM(S): Mus musculus

PROVIDER: GSE296378 | GEO | 2026/03/29

REPOSITORIES: GEO

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Project description:Background: Epigenetics is involved in various human diseases. Smoking is one of the most common environmental factors causing epigenetic changes. The DNA methylation changes and mechanisms after quitting smoking have not yet been defined. The present study examined the changes in DNA methylation level before and after short-term smoking cessation and explored the potential mechanism. Methods: Whole blood and clinical data were collected in 8 patients before and after short-term smoking cessation, DNA methylation was assessed, and differentially methylated sites were analyzed, followed by a comprehensive analysis of the differentially methylated sites with clinical data. GO/KEGG enrichment and protein-protein interaction (PPI) network identified the hub genes. The differentially methylated sites were detected by GEO2R between former smoking and current smoking in GSE50660 from the GEO database. Then, a Venn analysis was carried out using the differentially methylated sites. GO/KEGG enrichment analysis was performed on the genes corresponding to the common DNA methylation sites, the PPI network was constructed, and hub genes were predicted. The enriched genes associated with the cell cycle were selected, and the gene expression was analyzed in pan-cancer based on the TCGA database. Results: Most of the DNA methylation levels were decreased after short-term smoking cessation; a total of 694 hypermethylated CPG sites and 3184 hypomethylated CPG sites were identified. The DNA methylation levels altered according to the clinical data (body weight, expiratory, and tobacco dependence score). Enrichment analysis, construction of PPI network, and pan-cancer analysis suggested that smoking cessation may be involved in various biological processes. Conclusions: Smoking cessation leads to epigenetic changes, mainly observed in the decline of most DNA methylation levels. Bioinformatics further identified the biologically relevant changes after short-term smoking cessation.

Dataset Information

Regulatory Effects of Smoking Cessation on the Cellular Microenvironment and Differentially Expressed Genes in Precancerous Lesions of Pulmonary Nodules in Mice based on Single-Cell RNA-Sequencing and Immune Repertoire-Sequencing

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