Figure 7.
miR-216a-dependent transcriptional changes in endothelial cells
RNA sequencing was performed to assess the transcriptomic changes in endothelial cells treated with scrambled or miR-216a precursor molecules. (A) Heatmap of the top differentially expressed genes in endothelial cells treated with precursor-miR-216a compared with endothelial cells treated with scrambled precursor showing log2 FPKM (color scale) values of dysregulated genes, with yellow and blue colors representing increased and decreased expression, respectively. (B) Number of differentially expressed genes enriched in GO terms and (C) KEGG pathway analysis of differentially expressed genes. N refers to the number of genes involved in the pathway and DE refers to the genes in our dataset of differently expressed genes, therefore DE/N stands for the gene ratio and P.DE the p value for over-representation of the GO/KEGG term in the set. (D and E) Real-time PCR analysis of the expression of five representative (D) downregulated and (E) upregulated genes, n = 3 independent experiments. Numerical data are presented as mean (error bars show SEM); statistical significance was calculated using two-tailed unpaired t test. ∗p < 0.05 vs. corresponding control group. (F and G) Real-time PCR analysis of the expression of five representative (F) downregulated and (G) upregulated genes in myocardial tissue of WT and KO mice, n refers to number of hearts (n = 4 for each condition). Model depicting the regulatory mechanisms by which miR-216a controls physiological cardiac angiogenesis levels and regulates cardiac function. Decreased mir-216a expression levels in (cardiac) endothelial cells not only increases autophagic activity through direct upregulation of PTEN/BECN1 and indirectly ATG5 expression with subsequent impairment of angiogenesis, but also majorly impacts on apoptosis signaling and cardiac inflammatory response, leading to cardiac pathologic remodeling, susceptibility to cardiac stress, and ultimately to cardiac dysfunction and heart failure.