Identification of senescence-related genes for potential therapeutic biomarkers of atrial fibrillation by bioinformatics, human histological validation, and molecular docking

Figures

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  Fig. 1. Flowchart of the research of identification of senescence-related genes for potential biomarkers of atrial fibrillation.

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  Fig. 2. DEGs and functional enrichment analysis of genes common to senescence-related genes. (A) Rows represent DEGs, and columns represent samples. DEGs that are upregulated or downregulated are represented by red and blue, respectively. (B) Upregulated DEGs are represented by red plot points, while downregulated DEGs are represented by blue plot points. (C) A venn diagram illustrating 7 overlapping genes in DEGs and senescence-related genes. (D) Differential expression levels of 7 common genes between SR and AF. (E) Analysis of 7 common genes using Gene Ontology (GO). Biological processes (BP) are represented by the red bars, cellular components (CC) by the yellow bars, and molecular functions (MF) by the blue bars. DEG, differentially expressed gene; AF, atrial fibrillation; SR, sinus rhythm.

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  Fig. 3. Weighted gene co-expression network analysis. (A) Sample dendrogram. (B) Process for selecting soft thresholds. (C) Cluster dendrogram. Co-expression modules are represented by specific colors. (D) Different gene co-expression modules adjacency heatmap. (E) Heatmap showing the correlation between AF and module eigengenes.

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  Fig. 4. The selection and enrichment analysis of hub genes. The top 30 hub genes in purple module (A), turquoise module (C) and blue module (E). Gene Ontology (GO) analysis of hub genes in purple module (B), turquoise module (D) and blue module (F). Biological processes (BP) are represented by the red bars, cellular components (CC) by the yellow bars, and molecular functions (MF) by the blue bars.

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  Fig. 5. Identification of ETS1, SP1 and WT1 and correlation with cardiovascular diseases. (A) Venn diagram showing 3 overlapping genes (ETS1, SP1, WT1) in DEGs and hub genes of three modules. (B) Relationship to cardiovascular diseases related to ETS1, SP1 and WT1 based on the CTD database. ETS1 are represented by the red bars, SP1 by the yellow bars, and WT1 by the blue bars. * Evidence of a marker in the disease.

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  Fig. 6. Protein–protein interaction network analysis of predicted target genes. Protein–protein interaction network analysis of predicted target genes of ETS1 (A), SP1 (B) and WT1 (C).

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  Fig. 7. Validation of ETS1, SP1, and WT1 in human LAA and in vitro and in vivo experiments. Representative immunohistochemical staining images of ETS1, SP1and WT1 (A) and quantification (B). Data are expressed as mean ± SD (n = 5 per group). scale bar = 100 μm. (C). Relative mRNA expression levels of ETS1, SP1 and WT1 in human LAA. Data are expressed as mean ± SD (n = 8 per group). (D). Western blot analyses of ETS1, SP1 and WT1 in human LAA and statistical plot. Data are expressed as mean ± SD (n = 6 per group) (E). Relative mRNA expression levels of ETS1, SP1 and WT1 in HL-1 cells treated with saline or Ang II. Data are expressed as mean ± SD (n = 8 per group). (F). Western blot analyses of ETS1, SP1 and WT1 in HL-1 cells treated with saline or Ang II and statistical plot. Data are expressed as mean ± SD (n = 6 per group). (G). Relative mRNA expression levels of ETS1, SP1 and WT1 in LAA of mice administered with saline or Ang II for 4 weeks. Data are expressed as mean ± SD (n = 6 per group). (H). Western blot analyses of ETS1, SP1 and WT1 in LAA of mice administered with saline or Ang II for 4 weeks. Data are expressed as mean ± SD (n = 6 per group).

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  Fig. 8. Molecular docking results of Navitoclax interaction with ETS1, SP1 and WT1. Molecular docking conformation of Navitoclax interaction with ETS1 (A), SP1 (B) and WT1 (C). (D). The hydrogen bonding sites and lengths formed by the interaction between Navitoclax and ETS1, SP1, WT1. (E). Navitoclax's affinity values for ETS1, SP1, and WT1.