Identification of PGF+ endothelial cells associated with plaque instability in carotid atherosclerosis by scRNA-seq and RNA-seq analysis.
This study identifies pro-inflammatory endothelial cell subsets associated with plaque instability in atherosclerosis, highlighting the complexity of endothelial cell roles in vascular health.
Where it sits
this study against the rest of the ara 290 corpusSummary and findings
This study analyzed endothelial cell heterogeneity in carotid atherosclerosis using single-cell RNA sequencing and bulk RNA sequencing. It identified three endothelial cell subsets significantly enriched in unstable plaques, associated with pro-inflammatory cytokine upregulation. Findings were validated in an ApoE-/- mouse model showing increased placental growth factor expression.
Abstract
Vascular endothelial dysfunction plays a critical role in the development of atherosclerosis; however, the mechanisms by which endothelial cells contribute to plaque instability remain incompletely understood. In this study, we performed an integrated analysis of single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq data to characterize endothelial cell heterogeneity associated with carotid plaque instability. Clustering analysis, gene set variation analysis (GSVA), differential gene expression analysis, and KEGG pathway enrichment were conducted to identify key endothelial cell subsets and their functional characteristics. We identified three endothelial cell subsets (subsets 9, 10, and 11) that were significantly enriched in unstable plaques and exhibited upregulation of multiple pro-inflammatory cytokines. Pathway analysis revealed that these subsets were associated with activation of the PI3K - Akt signaling pathway and other inflammation-related pathways. Furthermore, findings were validated in an apolipoprotein E-deficient (ApoE-/-) mouse model, where increased expression of placental growth factor (PGF) and a higher proportion of PGF-positive endothelial cells were observed in atherosclerotic lesions. In conclusion, this study reveals the heterogeneity of endothelial cells in atherosclerotic plaques and identifies pro-inflammatory endothelial subsets potentially associated with plaque instability, providing new insights into the pathogenesis of atherosclerosis.<b>Trial registration</b>: This study was approved by the Experimental Animal Ethics Committee of Guizhou University of Chinese Medicine (approval number: 2,024,010).
Background
This paper addresses the role of vascular endothelial dysfunction in atherosclerosis, particularly how endothelial cells contribute to plaque instability. Prior research has indicated that endothelial cell heterogeneity may influence atherosclerotic progression, but specific mechanisms remain unclear. Understanding these mechanisms could provide insights into potential therapeutic targets for atherosclerosis management.
Methods
The study utilized single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing to analyze endothelial cell populations associated with carotid plaque instability. The population studied included endothelial cells from atherosclerotic plaques, with a focus on identifying key subsets. The analysis involved clustering, gene set variation analysis (GSVA), differential gene expression analysis, and KEGG pathway enrichment.
Results
The study identified three endothelial cell subsets (subsets 9, 10, and 11) that were significantly enriched in unstable plaques. These subsets exhibited upregulation of multiple pro-inflammatory cytokines and were associated with the activation of the PI3K - Akt signaling pathway. In the ApoE-/- mouse model, increased expression of placental growth factor (PGF) and a higher proportion of PGF-positive endothelial cells were observed in atherosclerotic lesions.
Interpretation
The findings contribute to the understanding of endothelial cell heterogeneity in atherosclerosis and suggest a link between pro-inflammatory endothelial subsets and plaque instability. However, the effect sizes and clinical significance of these findings remain unclear, particularly given the reliance on animal models. The study's conclusions may be limited by the lack of direct human evidence and the potential for confounding factors in the mouse model.
Key findings
- Three endothelial cell subsets (subsets 9, 10, and 11) significantly enriched in unstable plaques.
- Upregulation of multiple pro-inflammatory cytokines in identified endothelial subsets.
- Activation of the PI3K - Akt signaling pathway in these subsets.
- Increased expression of placental growth factor (PGF) in atherosclerotic lesions in ApoE-/- mice.
- Higher proportion of PGF-positive endothelial cells observed in atherosclerotic lesions.
Limitations
- Findings based on mouse models, not directly applicable to humans.
- Correlational data does not establish causation.
- Single-site study may limit generalizability.
- Short follow-up period in animal model.