miR-208b Reduces the Expression of Kcnj5 in a Cardiomyocyte Cell Line

• Main Authors: Julia Hupfeld, Maximilian Ernst, Maria Knyrim, Stephanie Binas, Udo Kloeckner, Sindy Rabe, Katja Quarch, Danny Misiak, Matthew Fuszard, Claudia Grossmann, Michael Gekle, Barbara Schreier
• Format: Article
• Language: English
• Published: MDPI AG 2021-06-01
• Series: Biomedicines
• Subjects: miR-208b; cardiomyocytes; Kcnj5; Ran; Rock2; heart
• Online Access: https://www.mdpi.com/2227-9059/9/7/719

MicroRNAs (miRs) contribute to different aspects of cardiovascular pathology, among them cardiac hypertrophy and atrial fibrillation. Cardiac miR expression was analyzed in a mouse model with structural and electrical remodeling. Next-generation sequencing revealed that miR-208b-3p was ~25-fold upregulated. Therefore, the aim of our study was to evaluate the impact of miR-208b on cardiac protein expression. First, an undirected approach comparing whole RNA sequencing data to miR-walk 2.0 miR-208b 3′-UTR targets revealed 58 potential targets of miR-208b being regulated. We were able to show that miR-208b mimics bind to the 3′ untranslated region (UTR) of voltage-gated calcium channel subunit alpha1 C and Kcnj5, two predicted targets of miR-208b. Additionally, we demonstrated that miR-208b mimics reduce GIRK1/4 channel-dependent thallium ion flux in HL-1 cells. In a second undirected approach we performed mass spectrometry to identify the potential targets of miR-208b. We identified 40 potential targets by comparison to miR-walk 2.0 3′-UTR, 5′-UTR and CDS targets. Among those targets, Rock2 and Ran were upregulated in Western blots of HL-1 cells by miR-208b mimics. In summary, miR-208b targets the mRNAs of proteins involved in the generation of cardiac excitation and propagation, as well as of proteins involved in RNA translocation (Ran) and cardiac hypertrophic response (Rock2).