Single-Cell Atlas of Endothelial Cells in Atherosclerosis: Identifying C1 CXCL12+ ECs as Key Proliferative Drivers for Immunological Precision Therapeutics in Atherosclerosis

Zhenzhen Zhao¹Yujiang Dong²Zhijie Zhao³Xiahou Zhikai⁴Cong Sun^2*

• ¹First College of Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China
• ²Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China
• ³Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
• ⁴China Institute of Sport and Health Science, Beijing Sport University, Beijing, China

Background: Atherosclerosis (AS) is a chronic inflammatory disease characterized by endothelial dysfunction, monocyte infiltration, smooth muscle proliferation, and extracellular matrix accumulation. Endothelial cell (EC) dysfunction plays a pivotal role in the initiation and progression of AS. Despite progress in traditional research methods, the complexity of cellular heterogeneity within the disease remains poorly understood, necessitating a more refined approach for uncovering disease mechanisms.Methods: In this study, we employed single-cell RNA sequencing (scRNA-seq) to map the endothelial cell landscape in AS comprehensively. By analyzing cellular heterogeneity, differentiation trajectories, and functional states, we identified critical endothelial subpopulations and their roles in the progression of AS. Functional enrichment and differentiation analyses were conducted, and the findings were validated through in vitro experiments.Results: The single-cell analysis revealed distinct EC subpopulations with unique contributions to AS progression. Among these, C1 CXCL12+ ECs emerged as a key subpopulation associated with endothelial differentiation, vascular remodeling, and inflammation. These cells demonstrated high proliferative potential and were enriched in pathways related to endothelial migration and repair. Through CCK-8, Transwell assay, EdU staining and angiogenesis ability, we found that knockdown of FOXM1 in C1 CXCL12+ ECs resulted in decreased proliferation, migration and invasion. Thus, it affects the progression of AS.Conclusion: This study provides a detailed single-cell atlas of endothelial cells in AS, identifying critical subpopulations, regulatory pathways, and key factors driving disease progression. The application of single-cell technologies paves the way for advancing our understanding of cardiovascular diseases and offers significant potential for developing personalized therapeutic strategies in immunology and precision medicine.