Single-cell dissection of cholesterol metabolism reveals cell-type-specific genes and hypercholesterolemia-associated...

This study was aimed at systematically characterizing cell-type-specific cholesterol metabolism in human blood samples and investigating the impact of hypercholesterolemia on the composition and gene expression of immune and stromal populations. Single-cell RNA sequencing (scRNA-seq) data from 31 human blood samples, encompassing 472,204 high-quality circulating cells, were analyzed. Pathway activities were assessed via gene set variation analysis (GSVA), and cell-type-specific markers were identified using differential expression analysis. Clinical cholesterol levels were used to stratify samples for comparative analysis of cell abundance and transcriptional changes. Monocytes and macrophages were found to exhibit heightened activity in cholesterol transport and dynamic response pathways. Five monocyte-enriched cholesterol metabolism genes (colony-stimulating factor 3 receptor, formyl peptide receptor 1, myeloid cell nuclear differentiation antigen, macrophage-expressed gene 1, and versican) were identified. Hypercholesterolemia was associated with increased abundance of macrophages, T cells, and natural killer cells, as well as decreased B cells and monocytes. Significant upregulation of proinflammatory genes such as VCAM1 in high-cholesterol conditions was revealed by differential gene expression analysis. Our study provided a comprehensive cell-type-specific atlas of cholesterol metabolism in human blood cells, highlighting metabolic heterogeneity and hypercholesterolemia-induced immune remodeling in the circulation. This resource offered novel insights into cholesterol-mediated immunomodulation.NEW & NOTEWORTHY Monocytes and macrophages display elevated activity in cholesterol transport and dynamic response pathways. Five monocyte-enriched genes (CSF3R, FPR1, MNDA, MPEG1, VCAN) are implicated in cholesterol metabolism regulation. Hypercholesterolemia reshapes circulating immune cell composition and upregulates proinflammatory genes (e.g., VCAM1), mediating immunomodulatory effects.