Plasma cfRNA was associated with calcification in carotid atherosclerotic plaques and may reflect aspects of underlying plaque biology and require validation in larger, independent populations.
Abstract
Vascular calcification (VC) is a hallmark of advanced atherosclerotic plaque biology. Circulating cell-free RNA (cfRNA) provides a non-invasive window into tissue transcriptional activity and may reflect plaque composition. We examined whether plasma cfRNA profiles are associated with carotid plaque calcification. Plasma cfRNA was profiled by RNA sequencing in 333 patients undergoing carotid endarterectomy, split into discovery (
n
= 216) and internal validation (
n
= 117) cohorts. Plaque calcification was quantified histologically. Differential expression analysis was performed using generalized linear models, followed by pathway enrichment. A cfRNA-derived gene score was constructed, and its incremental value beyond clinical risk factors was evaluated. Cellular deconvolution was applied to explore potential cfRNA origins. In this exploratory analysis, 13 genes showed nominal association with calcification in the discovery cohort (
p
< 0.001). Of these, 11 (84.6%) demonstrated concordant directionality in validation (Spearman
ρ
= 0.71,
p
= 0.008). Pathway analysis suggested potential involvement of oxidative phosphorylation and calcium signaling. The 11-gene score remained associated with calcification after adjustment for clinical risk factors and improved model performance. Plasma cfRNA was associated with calcification in carotid atherosclerotic plaques and may reflect aspects of underlying plaque biology. These findings are exploratory and require validation in larger, independent populations.
Background Carotid plaque calcification is an active multicellular process with heterogeneous clinical implications. However, endothelial cell (EC) heterogeneity and plaque-region-specific EC states associated with calcified lesions remain incompletely characterized. Methods We performed an exploratory integrative analysis of the public single-cell RNA sequencing dataset GSE159677, comprising paired calcified core (AC) and proximal adjacent (PA) tissues from three patients, together with a single-center proteomic cohort of three additional patients with paired AC and PA samples. Major plaque cell populations and EC subclusters were identified by unsupervised clustering and canonical markers. Calcium signaling activity, pathway enrichment, ligand–receptor communication, and Monocle2 pseudotime trajectories were analyzed. Transcriptomic findings were compared with differentially expressed proteins to identify cross-omics candidate molecules. Results A total of 35,890 cells were classified into seven major cell types. AC and PA tissues showed distinct cellular compositions and signaling patterns. Re-clustering of 4,925 ECs identified six subclusters, including a calcium signaling-high EC cluster enriched for extracellular matrix organization, inflammatory signaling, cytoskeletal regulation, and endothelial-to-mesenchymal transition-related programs. CellChat analysis indicated plaque-region-specific communication networks involving ECs, immune cells, fibroblasts, and smooth muscle cells. Pseudotime analysis suggested heterogeneous EC state transitions rather than a definitive longitudinal progression. Cross-omics comparison identified eight candidate molecules, FABP4, FABP5, MYL12A, POSTN, S100A10, SERPINB1, SOD2, and TMSB10, with concordant changes across transcriptomic and preliminary proteomic analyses. Conclusion These exploratory findings characterize plaque-region-specific EC heterogeneity associated with carotid plaque calcification and nominate candidate pathways and molecules for further validation in larger cohorts and functional models.
Yi-Cong Zhou, Dan-Dan Lin, Yan Yan et al.· International Journal of Gen...· 0 citations
BACKGROUND AND AIMS
Despite effective LDL-cholesterol reduction, residual cardiovascular risk persists. We investigated whether a specific fatty-acid phenotype predicts structural plaque instability and cardiovascular events independent of lipoprotein burden.
METHODS
This multi-cohort study integrated a clinical discovery cohort (n = 201, GC-MS profiling) with the UK Biobank validation cohort (n = 263,481, NMR profiling). Unsupervised clustering derived fatty-acid phenotypes. Structural vulnerability was assessed via optical coherence tomography (OCT) in 138 patients. The primary endpoint was MACE (cardiovascular death, non-fatal MI, stroke).
RESULTS
Two phenotypes emerged: Phenotype B, characterized by omega-3 depletion. Paradoxically, despite lower LDL-cholesterol (2.7 vs 3.0 mmol/L; p = 0.050) and triglycerides (1.5 vs 2.0 mmol/L; p = 0.041), Phenotype B exhibited significantly thinner fibrous caps (68.0 vs 94.2 μm; p = 0.003) and higher thin-cap fibroatheroma prevalence (49.0% vs 20.0%; p = 0.002) on OCT. In the UK Biobank, Phenotype B was associated with increased MACE risk in participants aged >60 years (HR 1.16; 95% CI 1.12-1.21; p < 0.001). In fully adjusted models, the hazard ratio was 1.12 (95% CI 1.08-1.16; p < 0.001), with 72% of excess risk mediated through glycemic, inflammatory, and adiposity pathways.
CONCLUSIONS
A latent fatty acid phenotype defined by omega-3 depletion and elevated omega-6/omega-3 ratio was associated with structural plaque vulnerability despite favorable atherogenic lipoprotein levels. In the UK Biobank, this phenotype was independently associated with increased MACE, most pronounced in participants aged 60 years and older. These observational findings warrant prospective validation.
Yuan Wang, Chaohui Han, Jiahui Yao et al.· Atherosclerosis· 0 citations
DNA methylation-based signatures were associated with incident ASCVD and modestly improved risk prediction beyond that of traditional risk factors, and an agnostic probe reliability-based approach was developed.
A. Barad, D. Khodasevich, P. F. Kho et al.· medRxiv· 0 citations
