Myocardial RNA Sequencing Reveals New Potential Therapeutic Targets in Heart Failure with Preserved Ejection Fraction

Figures

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  Figure 1. Gene expression differences between HFpEF and Control groups. (A) Bar plot showing the proportion of uniquely mapped, multiple mapped, and unmapped reads. The x-axis identifies the samples, and the y-axis shows the number of reads per million. (B) Bar plot comparing the relative abundance of each biotype (y-axis) in both sample types (x-axis). Green: Control; Red: HFpEF. (C) Distribution of read counts (log2) per sample (A) before normalization and (B) after normalization—HSKG normalization and RIN adjustment. Green: Control; Red: HFpEF. (D) Principal component analysis (PCA) of the mRNA from cardiac biopsy samples. Green: Control; Red: HFpEF. (E) Hierarchical clustering dendrogram using the Euclidean method for the 10 samples. Green: Control; Red: HFpEF.

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  Figure 2. Functional classification of the DEGs. (A) Density plot showing the number of differentially expressed genes between HFpEF and Control groups with a false discovery rate (FDR) adjusted p-value < 0.05. The x-axis represents significantly dysregulated genes, and the y-axis shows the density of the log2 fold change. Downregulated genes are represented in red and upregulated genes in green. (B) GO biological process (BP) enrichment analysis, (C) GO molecular function (MF) enrichment analysis, and (D) GO cellular component (CC) enrichment analysis of the 306 significant DEGs from HFpEF vs. Control. The color of the dots indicates significance; the size of the dots indicates the gene count for each category; and the gene ratio indicates the enrichment level.

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  Figure 3. microRNA expression differences between HFpEF and Control groups. (A) Percentage of the reads mapped to different types of RNA and unaligned reads. (B) Distribution of the read counts (log2) per sample (A) before normalization and (B) after normalization—HSKG normalization and RIN adjustment. Green: Control; Red: HFpEF. (C) PCA score plot of the microRNA samples. Green and red colors represent the Control and HFpEF patients, respectively. (D) Hierarchical clustering dendrogram using the Euclidean method for the 10 microRNA samples. Green: Control; Red: HFpEF. (E) Summary of genes considered for microRNA–mRNA interaction analysis.

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  Figure 4. Correlation between each mRNA and corresponding miRNA(s). (A) Venn diagram of the number of miRNA–mRNA negative correlated pairs. (A) represents the number of pairs with correlation < 0 and padj < 0.05, and (B) represents the number of targets (miRNA–mRNA pairs) in public databases, which are present in our full dataset. (B) Network of miRNA–mRNA pairs with significant negative correlations (padj < 0.05). Circles represent miRNAs and squares mRNAs. Red fill means upregulated miRNA/mRNA, while green fill means downregulated miRNA/mRNA. Arrows indicate miRNA–mRNA pairs. Arrow color represents the score of the interaction; in this case, only weak-to-strong negative interactions (red arrows) are represented. Arrow width is proportional to the number of appearances in the database.

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  Figure 5. Relative mRNA expression of HAPLN1 and NPPB genes in human cardiac myocyte line transfected with hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-140-3p, or hsa-miR-4429. Results represent the mean ± SD of five replicates. Unpaired Student’s t-test was applied to compare the differences between conditions. Results were considered statistically significant when ** p < 0.01 and *** p < 0.001.