Circular Network of Coregulated Sphingolipids Dictates Chronic Hypoxia Damage in Patients With Tetralogy of Fallot

Figures

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  Figure 1. Workflow of the sphingolipid metabolism analysis of this study.

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  Figure 2. Identification of sphingolipid metabolism in Tetralogy of Fallot (TOF). (A) Lipid metabolic process was significantly enriched in the GSE132176 dataset by the gene set enrichment analysis (GSEA). (B) Lipid metabolic processes were significantly enriched in the GSE141955 dataset by the GSEA. (C,D) Sphingolipid metabolism was significantly downregulated in TOF compared with atrial septal defects (ASDs) by the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome analyses, while sphingolipid transfer activity and the sphingolipid biosynthetic process were not significantly different. (E,F) Sphingolipid metabolism was significantly downregulated in TOF compared with healthy donors by the KEGG and Reactome analyses, while sphingolipid transfer activity and the sphingolipid biosynthetic process were not significantly different.

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  Figure 3. Clustering analysis of genes involved in sphingolipid metabolism and the sphingolipid biosynthetic process. (A,C) Heatmap showing the genes involved in sphingolipid metabolism and the sphingolipid biosynthetic process between the TOF and atrial septal defect (ASD) samples in the GSE132176 dataset. (B,D) Heatmap showing the genes involved in sphingolipid metabolism and the sphingolipid biosynthetic process between patients with TOF and healthy donors in the GSE141955 dataset.

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  Figure 4. Statistical analysis of differential metabolites between the TOF and ASD samples. (A,B) Volcano plot showing the differential metabolites between the TOF and ASD samples. Red and green indicate upregulated and downregulated metabolites in the TOF group, respectively. (C,D) OPLS-DA score plot. The X-axis represents the predicted component score, while the Y-axis represents the orthogonal component score. (E,F) Heatmap showing the differential metabolites of the right atrium (RA) and plasma between patients with TOF and ASD. (G,H) Plot of the RA and plasma OPLS-DA between patients with TOF and ASD.

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  Figure 5. Functional enrichment of the differential metabolites and differential analysis of sphingolipid metabolism-related metabolites. (A,B) Top 20 differential metabolites between the TOF and ASD samples. (C,D) The KEGG pathway enrichment of the differential metabolites between the TOF and ASD samples. (E) The common differential metabolites of plasma between our metabolites and the sphingomyelin metabolite library. (F) Heatmap of common differential metabolites of plasma between our metabolites and the sphingomyelin metabolite library. (G) The common differential metabolites of RA tissue between our metabolites and the sphingomyelin metabolite library. (H) Heatmap of common differential metabolites of RA tissue between our metabolites and the sphingomyelin metabolite library.

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  Figure 6. Association analysis between differential sphingomyelin metabolites and important clinical phenotypes in TOF. (A,B) Heatmaps based on z-scores of differential sphingomyelin metabolites and clinically associated phenotypes. (C,D) Correlation analysis between z-scores of differential sphingomyelin metabolites and clinically important phenotypes in TOF.

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  Figure 7. Identification of core regulatory genes related to sphingomyelin metabolism in TOF. (A) Common genes among the GES141955 differentially expressed genes (DEGs), GSE132176 DEGs, GES141955 consensus genes, GES132176 consensus genes, and sphingomyelin metabolism-related genes. (B) Gene-pathway interaction network by partial correlation analysis based on standardized gene expression values and sphingomyelin metabolic pathway scores in the GSE141955 dataset. (C) Gene-pathway interaction network by partial correlation analysis based on standardized gene expression values and sphingomyelin metabolic pathway scores in the GSE132176 dataset. (D,E) Expression of core genes in GSE132176 and GSE141955. (F,G) QT-PCR validation of the expression of the four core genes. (H) Mechanism plot of the study. #p < 0.05.