Immune cell-specific smoking-related expression characteristics are revealed by re-analysis of transcriptomes from the CEDAR cohort

Introduction
Smoking is known to affect whole-blood expression and methylation profiles. Although whole-genome methylation studies indicated that effects observed in blood may be driven by changes within leukocyte subtypes, these phenomena have not been explored using expression profiling.

Material and methods
This study reanalyzed data from the Correlated Expression and Disease Association Research (CEDAR) patient cohort recruited by Momozawa et al. (E-MTAB-6667). Data from gene expression profiling of immunomagnetically sorted CD4⁺, CD8⁺, CD14⁺, CD15⁺, and CD19⁺ cells were processed. Differential expression analyses were conducted in each immune cell type, followed by gene ontology analysis and supplementary investigations.

Results
Ninety-four differentially expressed genes were found (CD8⁺ n = 58, CD14⁺ n = 20, CD4⁺ n = 14, CD19+ n = 2). Two key smoking-related genes were overexpressed in specific cell types: LRRN3 (CD4⁺, CD8⁺) and MMP25 (CD8⁺, CD14⁺). In CD4+ cells smoking was associated with reduced expression of the NK cell receptor KLRB1, suggesting CD4⁺ subpopulation shifts and differences in interferon signaling (reduced IRF1 and IL18RAP in smokers). Key results and their integration with an immune protein-protein interaction network revealed that smoking influences integrins in CD8⁺ cells (ITGB7, ITGAL, ITGAM, ITGB2). C-type lectin CLEC4A was reduced in CD8⁺ cells and CLEC10A was increased in CD14⁺ cells from smokers; moreover, CLEC5A (CD8⁺), CLEC7A (CD8⁺) and CLEC9A (CD19⁺) were related to smoking in supplementary analyses. CD14⁺ cells from smokers exhibited overexpression of LDLR and the formyl peptide receptor FPR3.

Conclusions
Smoking specifically alters vital immune regulation genes in lymphocyte subtypes, especially CD4⁺, CD8⁺ and CD14⁺ cells.