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Updated: Jun 27, 2025
Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus
Published on: December 7, 2013
Unveiling Atherosclerotic Plaque Heterogeneity and SPP1+/VCAN+ Macrophage Subtype Prognostic Significance Through
Xiang Xu1,2, Fuling Qiu2, Man Yang3
1School of Medicine, Yunnan University, Kunming City, Yunnan Province, People's Republic of China.
Background:
Dysregulated macrophages are important causes of Atherosclerosis (AS) formation and increased plaque instability, but the heterogeneity of these plaques and the role of macrophage subtypes in plaque instability have yet to be clarified.
Methods:
This study integrates single-cell and bulk-seq data to analyze atherosclerotic plaques. Unsupervised clustering was used to reveal distinct plaque subtypes, while survival analysis and gene set variation analysis (GSVA) methods helped in understanding their clinical outcomes. Enrichment of differential expression of macrophage genes (DEMGs) score and pseudo-trajectory analysis were utilized to explore the biological functions and differentiation stages of macrophage subtypes in AS progression. Additionally, CellChat and the BayesPrism deconvolution method were used to elucidate macrophage subtype interaction and their prognostic significance at single-cell resolution. Finally, the expression of biomarkers was validated in mouse experiments.
Results:
Three distinct AS plaque subtypes were identified, with cluster 3 plaque subtype being particularly associated with higher immune infiltration and poorer prognosis. The DEMGs score exhibited a significant elevation in three macrophage subtypes (SPP1+/VCAN+ macrophages, IL1B+ macrophages, and FLT3LG+ macrophages), associated with cluster 3 plaque subtype and highlighted the prognostic significance of these subtypes. Activation trajectory of the macrophage subtypes is divided into three states (Pre-branch, Cell fate 1, and Cell fate 2), and Cell fate 2 (SPP1+/VCAN+ macrophages, IL1B+ macrophages, and FLT3LG+ macrophages dominant) exhibiting the highest DEMGs score, distinct interactions with other cell components, and relating to poorer prognosis of ischemic events. This study also uncovered a unique SPP1+/VCAN+ macrophage subtype, rare in quantity but significant in influencing AS progression. Machine learning algorithms identified 10 biomarkers crucial for AS diagnosis. The validation of these biomarkers was performed using Mendelian Randomization analysis and in vitro methods, supporting their relevance in AS pathology.
Conclusion:
Our study provides a comprehensive view of AS plaque heterogeneity and the prognostic significance of macrophage subtypes in plaque instability.
Insights
This study reveals distinct atherosclerosis plaque subtypes and identifies specific macrophage subtypes (SPP1+/VCAN+, IL1B+, FLT3LG+) linked to poor prognosis and increased immune infiltration, offering new diagnostic biomarkers.
Area of Science:
- Cardiovascular Biology
- Immunology
- Genomics
Background:
- Dysregulated macrophages contribute significantly to atherosclerosis (AS) development and plaque instability.
- The intricate heterogeneity of atherosclerotic plaques and the precise roles of macrophage subtypes in driving plaque instability remain incompletely understood.
Purpose of the Study:
- To comprehensively analyze atherosclerotic plaque heterogeneity using integrated single-cell and bulk sequencing data.
- To elucidate the functional roles and prognostic significance of diverse macrophage subtypes in AS progression and plaque instability.
Main Methods:
- Integrated single-cell and bulk sequencing data analysis.
- Unsupervised clustering for plaque subtype identification, survival analysis, and gene set variation analysis (GSVA).
- Differential expression of macrophage genes (DEMGs) score, pseudo-trajectory analysis, CellChat, and BayesPrism for cell interaction and differentiation analysis.
Main Results:
- Identified three distinct AS plaque subtypes, with cluster 3 exhibiting high immune infiltration and poor prognosis.
- Elevated DEMGs scores in SPP1+/VCAN+, IL1B+, and FLT3LG+ macrophage subtypes correlated with cluster 3 and poorer outcomes.
- Macrophage differentiation trajectories revealed a 'Cell fate 2' state dominant in these subtypes, associated with adverse clinical events and distinct cellular interactions.
Conclusions:
- Provides a detailed map of AS plaque heterogeneity and the prognostic implications of specific macrophage subtypes.
- Highlights the clinical relevance of macrophage subtypes, particularly SPP1+/VCAN+ macrophages, in AS progression and plaque instability.
- Identified potential diagnostic biomarkers for AS through machine learning and validated their relevance.
