MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1.

Sadie C Slater, Eva Jover, Andrea Martello, Tijana Mitić, Iker Rodriguez-Arabaolaza, Rosa Vono, Valeria V Alvino, Simon C Satchell, Gaia Spinetti, Andrea Caporali, Paolo Madeddu

Molecular therapy : the journal of the American Society of Gene Therapy 2018 Dec 05

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MicroRNAs regulate endothelial function and angiogenesis, but their implication in pericyte biology remains undetermined. A PCR array, covering a panel of 379 human microRNAs, showed microRNA-532-5p to be one of the most differentially modulated by hypoxia, which was confirmed by qPCR in both skeletal muscle and adventitial pericytes. Furthermore, microRNA-532-5p was upregulated in murine muscular pericytes early after experimentally induced ischemia, decreasing below baseline after reperfusion. Transfection of human pericytes with anti-microRNA, microRNA-mimic, or controls indicates microRNA-532-5p modulates pro-angiogenic activity via transcriptional regulation of angiopoietin-1. Tie-2 blockade abrogated the ability of microRNA-532-5p-overexpressing pericytes to promote endothelial network formation in vitro. However, angiopoietin-1 is not a direct target of microRNA-532-5p. In silico analysis of microRNA-532-5p inhibitory targets associated with angiopoietin-1 transcription indicated three potential candidates, BACH1, HIF1AN, and EGLN1. Binding of microRNA-532-5p to the BACH1 3' UTR was confirmed by luciferase assay. MicroRNA-532-5p silencing increased BACH1, while a microRNA-532-5p mimic decreased expression. Silencing of BACH1 modulated angiopoietin-1 gene and protein expression. ChIP confirmed BACH1 transcriptional regulation of angiopoietin-1 promoter. Finally, microRNA-532-5p overexpression increased pericyte coverage in an in vivo Matrigel assay, suggesting its role in vascular maturation. This study provides a new mechanistic understanding of the transcriptional program orchestrating angiopoietin-1/Tie-2 signaling in human pericytes. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Citation

Sadie C Slater, Eva Jover, Andrea Martello, Tijana Mitić, Iker Rodriguez-Arabaolaza, Rosa Vono, Valeria V Alvino, Simon C Satchell, Gaia Spinetti, Andrea Caporali, Paolo Madeddu. MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1. Molecular therapy : the journal of the American Society of Gene Therapy. 2018 Dec 05;26(12):2823-2837