Cross-Disease Identification of FBP1 in Atherosclerosis and Calcific Aortic Valve Disease via Integrated Bioinformatics and Validation: Functional Analysis and Therapeutic Target Exploration

Figures

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  Figure 1. Overall design of the study. Significance: P < 0.05 (*), P < 0.0001 (****).

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  Figure 2. Cross-disease genes expression analysis. (A and B) Volcano plots of DEGs in AS and CAVD. (C and D) Heatmaps illustrate the expression patterns of corresponding DEGs in AS and CAVD. (E and F) Venn diagram illustrates the intersection analysis of CGs in AS and CAVD.

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  Figure 3. Integrated analysis of CGs and identification of metabolism-related CGs. (A and B) GO and KEGG enrichment analyses of 147 consensus CGs in AS and CAVD. (C) Venn diagram showing the intersection between shared DEGs and metabolism-related genes, identifying 10 overlapping candidates. (D and E) Co-expression correlations of the 10 hub genes in AS and CAVD datasets. (F and G) Expression levels of the 10 hub genes in AS and CAVD datasets. Significance: P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).

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  Figure 4. Identification of hub genes in AS and CAVD by machine learning. (A and B) LASSO regression analysis in the AS dataset. (C and D) Random forest (RF) analysis in the AS dataset. (E and F) LASSO regression in the CAVD dataset. (G and H) RF analysis in the CAVD dataset.

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  Figure 5. Identification and expression validation of candidate genes. (A) Intersection of LASSO and RF results across AS and CAVD cohorts identifying FBP1 and KRT18 as common candidate genes. (B–E) Expression profiles of candidate genes. Significance: P < 0.05 (*), P < 0.01 (**), P < 0.001 (***), P < 0.0001 (****); ns = not significant.

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  Figure 6. Diagnostic performance of the FBP1-based prediction model. (A–D) ROC curves demonstrating the predictive accuracy of FBP1 in AS and CAVD datasets. (E–H) Calibration curves showing agreement between predicted and actual outcomes in both diseases. (I–L) Decision curve analyses indicating clinical net benefit of the FBP1-based model in AS and CAVD. (M–N) Nomogram and schematic representations of AS and CAVD prediction models incorporating FBP1 expression.

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  Figure 7. Immune infiltration analysis and correlation with FBP1 expression. (A) Relative proportions of 22 immune-cell subsets in AS and controls. (B) Relative proportions of 22 immune-cell subsets in CAVD and controls. (C) Differential immune-cell abundances in AS vs controls. (D) Differential immune-cell abundances in CAVD vs controls. (E) Spearman correlations between FBP1 expression and immune-cell fractions in AS. (F) Spearman correlations between FBP1 expression and immune-cell fractions in CAVD. Significance: P < 0.05 (*), P < 0.01 (**), P < 0.001 (***); ns = not significant.

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  Figure 8. Pathway enrichment and VSMC phenotype coupling of FBP1. (A and B) GSEA of FBP1 in AS datasets. (C and D) GSEA of FBP1 in CAVD datasets. (E and F) FBP1–VSMC marker correlations in AS datasets (Spearman, P<0.05).

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  Figure 9. Regulatory landscape of FBP1. (A) Chromosomal location of FBP1. (B) Protein-protein interaction network showing FBP1-centered metabolic associations. (C) Venn diagram of transcription factors predicted by ENCODE, hTFtarget, and KnockTF. (D) Integrated regulatory network of FBP1 involving transcription factors and microRNAs.

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  Figure 10. Molecular docking analysis of candidate compounds targeting FBP1. (A–H) Docking results for chrysin, myricetin, kaempferol, and apigenin with FBP1.

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  Figure 11. HE and Oil Red O staining of atherosclerotic arteries from different sites. (A) Common carotid artery—less-affected intima (CCA-LA): HE, Scale bar: 2.5 mm; Oil Red O (ORO), Scale bar: 250 µm. Common carotid artery—advanced atherosclerotic intima (CCA-Adv): HE, Scale bar: 2.5 mm; ORO, Scale bar: 250 µm. (B) Abdominal aorta—healthy: HE, Scale bar: 2.5 mm; ORO, Scale bar: 250 µm. Femoral artery—atherosclerosis: HE, Scale bar: 2.5 mm; ORO, Scale bar: 250 µm.

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  Figure 12. Experimental validation of FBP1 expression in AS and CAVD tissues. (A and B) Western blot of FBP1 in lower-extremity arteriosclerosis obliterans (LE-ASO) vs normal controls (NC); densitometry. (C and D) FBP1 in common carotid artery—advanced atherosclerotic intima (CCA-Adv) vs less-affected intima (CCA-LA); densitometry. (E and F) FBP1 in CAVD vs NC; densitometry. (G) RT-qPCR of FBP1 mRNA in LE-ASO vs NC. (H) RT-qPCR in CCA-Adv vs CCA-LA. (I) RT-qPCR in CAVD vs NC. Data: mean ± SEM (n = 5 per group). Significance: P < 0.01 (**), P < 0.001 (***).