Tobacco smoking is associated with sex- and plaque-type specific upregulation of CRLF1 in atherosclerotic lesions

• Published in: Atherosclerosis 2024-10, Vol.397, p.118554, Article 118554
• Main Authors: Lan, Tian, Palm, Kaylin C.A., Hoeben, Luka, Diez Benavente, Ernest, Perry, R. Noah, Civelek, Mete, de Kleijn, Dominique P.V., den Ruijter, Hester M., Pasterkamp, Gerard, Mokry, Michal
• Format: Article
• Language: English
• Subjects: Aged; Carotid Artery Diseases - genetics; Carotid Artery Diseases - metabolism; Carotid Artery Diseases - pathology; Female; Gene Regulatory Networks; Humans; Male; Middle Aged; Plaque, Atherosclerotic; Receptors, Cytokine - genetics; Receptors, Cytokine - metabolism; Risk Factors; Sex Factors; Single-Cell Analysis; Smokers; Tobacco Smoking - adverse effects; Transcriptome; Up-Regulation
• ISSN: 0021-9150; 1879-1484; 1879-1484
• DOI: 10.1016/j.atherosclerosis.2024.118554

Tobacco smoking is a known risk factor for atherosclerotic disease, with more elevated risks in women compared to men. We hypothesized that atherosclerotic plaques from smokers show different gene expression patterns compared to non-smokers, in a sex-specific manner. Gene expression data of 625 carotid plaques (151 females and 474 males) were analyzed for differential gene expression between current smokers (n = 226) and non-smokers (n = 399). All analyses were stratified by sex and by molecular plaque characteristics. Finally, we projected the activity of gene regulatory networks and utilized single-cell transcriptomics from 38 plaques (26 males and 12 females) to interpret the sex- and plaque-type specific signals. We observed higher expression levels of CRLF1 gene in atherosclerotic plaques from smokers compared to non-smokers (log2FC = 0.48, FDR = 0.012). CRLF1 upregulation was interacting with sex (p = 0.01) and was more pronounced in females (log2FC = 0.93, p = 1.53E-05) compared to males (log2FC = 0.35, p = 0.0018). Through single-cell RNA-seq analysis, we identified the highest CRLF1 expression within the transitioning and synthetic smooth muscle cell populations. CRLF1 expression was increased in fibro-inflammatory and fibro-cellular plaque types. Gene annotations pointed to increased expression of CRLF1 in networks with extracellular matrix related genes. Atherosclerotic plaques from current smokers show sex-dependent upregulation of smooth muscle cell gene CRLF1. This may explain the different contributions of smoking to cardiovascular risk in females. [Display omitted] •Smoking is significantly associated with upregulation of CRLF1 gene expression in carotid plaque.•Female smokers exhibit significantly higher overexpression of CRLF1 gene compared to male smokers.•The expression of CRLF1 gene in carotid plaque is influenced by plaque composition and molecular type.•CRLF1 gene shows high expression in ACTA2+ smooth muscle cells.