miR-182-5p is an evolutionarily conserved Tbx5 effector that impacts cardiac development and electrical activity in zebrafish

Year: 2019

Authors: Guzzolino E., Pellegrino M., Ahuja N., Garrity D., D’Aurizio R., Groth M., Baumgart M., Hatcher C.J., Mercatanti A., Evangelista M., Ippolito C., Tognoni E., Fukuda R., Lionetti V., Pellegrini M., Cremisi F., L. Pitto L.

Autors Affiliation: Institute of Clinical Physiology, National Research Council, IFC, Pisa, Italy; National Institute of Optics, CNR, Pisa, Italy; Department of Biology, Colorado State University (CSU), Fort Collins, CO, USA; Institute of Informatics and Telematics, CNR, Pisa, Italy; The Leibniz Institute on Aging, Fritz Lipmann Institute (FLI), Jena, Germany; Department of Bio-Medical Sciences, Philadelphia College of Osteopathic Medicine, Philadelphia, PA, USA; Department of Clinical and Experimental Medicine, University of Pisa; Department of Developmental Genetics, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany; Institute of Life Sciences, Scuola Superiore Sant´Anna, Pisa, Italy; UOS Anesthesiology, Fondazione Toscana “”G.Monasterio””, Pisa, Italy ; Scuola Normale Superiore, Pisa, Italy

Abstract: To dissect the TBX5 regulatory circuit, we focused on microRNAs (miRNAs) that collectively contribute to make TBX5 a pivotal cardiac regulator. We profiled miRNAs in hearts isolated from wild-type, CRE, Tbx5(lox/+)and Tbx5(del/+) mice using a Next Generation Sequencing (NGS) approach. TBX5 deficiency in cardiomyocytes increased the expression of the miR-183 cluster family that is controlled by Kruppel-like factor 4, a transcription factor repressed by TBX5. MiR-182-5p, the most highly expressed miRNA of this family, was functionally analyzed in zebrafish. Transient overexpression of miR-182-5p affected heart morphology, calcium handling and the onset of arrhythmias as detected by ECG tracings. Accordingly, several calcium channel proteins identified as putative miR-182-5p targets were downregulated in miR-182-5p overexpressing hearts. In stable zebrafish transgenic lines, we demonstrated that selective miRNA-182-5p upregulation contributes to arrhythmias. Moreover, cardiac-specific down-regulation of miR-182-5p rescued cardiac defects in a zebrafish model of Holt-Oram syndrome. In conclusion, miR-182-5p exerts an evolutionarily conserved role as a TBX5 effector in the onset of cardiac propensity for arrhythmia, and constitutes a relevant target for mediating the relationship between TBX5, arrhythmia and heart development.


KeyWords: microRNAs, Holt-Oram syndrome, Zebrafish, Cardiac development
DOI: 10.1007/s00018-019-03343-7