PIK3R1, SPNB2, and CRYAB as Potential Biomarkers for Patients with Diabetes and Developing Acute Myocardial Infarction

Figures

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  Figure 1. Integrated bioinformatics analysis of AMI GEO datasets. (a) The normalization of three AMI datasets was applied through Log2 transformation. (b) Heatmap of three datasets demonstrated distinguished features between AMI and SHAM heart samples. (c) The gene cluster in PCA loading score. (d) The chord diagram of three AMI GEO datasets. (e) The KEGG topology enrichment analysis after integrated bioinformatics analysis. (f) The PPI topology analysis after integrated bioinformatics analysis. (g) The Gene Regulatory Networks TF-gene interactions analysis after integrated bioinformatics analysis.

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  Figure 2. Integrated bioinformatics analysis of Diabetes GEO datasets. (a) The normalization of three diabetes datasets was applied through Log2 transformation. (b) Heatmap of three datasets demonstrated distinguished features between diabetes and WT skeletal muscle samples. (c) The gene cluster in PCA loading score. (d) The PPI topology analysis after integrated bioinformatics analysis. (e) The Gene Regulatory Networks TF-gene interactions analysis after integrated bioinformatics analysis.

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  Figure 3. Differentially expressed transcripts among diabetes DEGs and AMI DEGs. (a) Venn diagram demonstrated 67 same expressed transcripts were screened among meta diabetes DEGs and meta AMI DEGs after integrated bioinformatics analysis. (b) GO slim of the 67 DEGs using WebGestalt. (c) The bubble plot of GO analysis about the 67 DEGs using Metascape. (d) The volcano plot of enrichment pathways after GO analysis using WebGestalt. (e) The KEGG topology enrichment analysis of the 67 DEGs using NetworkAnalyst. (f) The PPI topology analysis of the 67 DEGs using NetworkAnalyst.

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  Figure 4. miRNA-hub genes network. Venn diagram among predicted miRNAs which bind the 5′UTR, CDS, and 3′UTR of hub genes PIK3R1 (a), SPNB2 (b), or CRYAB (c) and miRNAs which were screened at 1d, 1w, and 8w after MI in GSE114695 for validation. (d) The network of screened hub genes and miRNAs after validation.

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  Figure 5. Validation of screened hub genes in mice MI model. The relative mRNA expression level of PIK3R1 (a), SPNB2 (b), and CRYAB (c) at 1d and 1w after MI using qPCR analysis. The protein expression levels of left ventricle at 1d and 1w after MI utilizing ELISA, including PIK3R1 (d), SPNB2 (e), and CRYAB (f). The expression levels of proteins, including PIK3R1 (g) and SPNB2 (h), were demonstrated to be negatively correlated to LVEF. The expression levels of protein CRYAB (i) were demonstrated to be positively correlated to LVEF. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns, not significant.

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  Figure 6. Validation of screened hub genes in human blood samples. (a) The protein expression levels of human blood samples between diabetes patients with and without AMI utilizing ELISA analysis, including PIK3R1, SPNB2, and CRYAB. (b) There is no significant difference in the hospital stay between diabetes patients with and without AMI. (c) The AUCs of three screened proteins of human blood samples between diabetes patients with and without AMI, including PIK3R1, SPNB2, and CRYAB, suggesting the three hub genes can be the diagnostic biomarkers for diabetic patients undergoing AMI. The protein expression levels of diabetic AMI patients' blood samples between before and after treatment utilizing ELISA analysis, including PIK3R1 (d), SPNB2 (e), and CRYAB (f). (g–h) There is no significant difference in the hospital stays between the diabetic AMI patients with high PIK3R1 and SPNB2 expression level and with low PIK3R1 and SPNB2 expression level. (i) The diabetic AMI patients with low PIK3R1 and SPNB2 expression level had the longer hospital stays compared to those with high PIK3R1 and SPNB2 expression level. (j) The AUCs of three screened proteins of diabetic AMI patients' blood samples…

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  Figure 7. The correlation analysis of the screened hub genes and clinical laboratory examination. The expression levels of proteins, including PIK3R1 (a) and SPNB2 (b), were demonstrated to be negatively correlated to CK expression, while the CRYAB (c) expression was demonstrated to be positively correlated to CK expression. The expression levels of proteins, including PIK3R1 (d) and SPNB2 (e), were demonstrated to be negatively correlated to CK-MB expression, while the CRYAB (f) expression was demonstrated to be positively correlated to CK-MB expression. The expression levels of proteins, including PIK3R1 (g) and SPNB2 (h), were demonstrated to be negatively correlated to cTnI expression, while the CRYAB (i) expression was demonstrated to be positively correlated to cTnI expression. The expression levels of proteins, including PIK3R1 (j) and SPNB2 (k), were demonstrated to be negatively correlated to AST expression, while the CRYAB (l) expression was demonstrated to be positively correlated to AST expression.