Multi-omics profiling of circulating proteins and metabolites in persistent atrial fibrillation: VDAC1 identified as a candidate circulating protein

Figures

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  Fig. 1. Study workflow. Plasma from 37 AF and 36 SVT patients was divided into a discovery cohort (n = 10 per group) for proteomics and metabolomics. An expanded cohort (n = 25 additional per group; 35 total per group) was used for ELISA. External support used one RNA-seq dataset (GSE282504), three microarray datasets, and single-nucleus RNA-seq data (GSE255612). Integrated multi-omic analysis identified VDAC1 as a candidate marker of AF-related mitochondrial injury.

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  Fig. 2. Proteomic analysis of plasma DEPs (n = 10 per group). (A) Volcano plot (red = upregulated, green = downregulated, gray = non-significant). (B) METASCAPE enrichment heatmap for upregulated DEPs. (C) METASCAPE enrichment heatmap for downregulated DEPs. (D) KEGG enrichment bubble plot. (E) VDAC1-centered PPI subnetwork.

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  Fig. 3. Metabolomic analysis (n = 10 per group). (A) Volcano plot. (B) Hierarchical clustering heatmap of metabolites. (C) Top 20 VIP metabolites. (D) KEGG enrichment bubble chart.

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  Fig. 4. Common KEGG pathways enriched in both proteomic and metabolomic data. Eight pathways met P < 0.01 in both datasets.

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  Fig. 5. ELISA validation of plasma VDAC1, CS, and SFRP2. (A) Full cohort (35 AF, 35 SVT). (B) Newly recruited patients only (25 AF, 25 SVT; discovery samples excluded). ***P < 0.001. Data are shown as median with interquartile range.

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  Fig. 6. VDAC1 expression in four GEO datasets. (A) GSE282504 (RNA-seq, 20 AF vs 10 SR). (B) GSE2240 (microarray, 10 AF vs 20 SR). (C) GSE41177 (microarray, 32 AF vs 6 SR). (D) GSE79768 (microarray, 14 AF vs 12 SR). *P < 0.05.

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  Fig. 7. Single-nucleus RNA-seq UMAP (GSE255612). (A) Cell subtypes. (B) UMAP of 15 cell types. (C) VDAC1 expression in AF and control cardiomyocytes.

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  Supplementary Figure S1. Quality control assessment of metabolomics data.(A) Pearson correlation analysis among QC samples. All QC samples showed r > 0.99.(B) Coefficient of variation (CV) distribution of all detected metabolites. An empirical cumulative distribution function (ECDF) plot is shown. Over 75% of metabolites had CV < 0.3.

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  Supplementary Figure S2. Metabolomic analysis (n = 10 per group). (A) PCA score plot. (B) OPLS-DA score plot.

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  Supplementary Figure S3. ROC curve for plasma VDAC1 in discriminating AF from SVT. AUC = 0.816 (95% CI: 0.715–0.917; P < 0.001).

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  Supplementary Figure S4. Proposed mechanism diagram. Left: AF cardiomyocyte (red = supported by previous studies and our data; blue dashed = speculative EV pathway). Right: SR cardiomyocyte with stable calcium homeostasis. SR, sinus rhythm; EVs, extracellular vesicles; ROS, reactive oxygen species.