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

Research output: Contribution to journalArticle

Abstract

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. Angiopoietin-1 is an important regulator of angiogenesis, yet little is known regarding its transcriptional regulation. Slater et al. (2018) demonstrate angiopoietin-1 induction in pericytes occurs via miR-532-5p regulation of BACH1 and modulation of this pathway promotes changes in endothelial permeability, vascular stability, and angiogenesis.

Original languageEnglish
Pages (from-to)2823-2837
Number of pages15
JournalMolecular Therapy
Volume26
Issue number12
DOIs
Publication statusPublished - Dec 5 2018

Fingerprint

Angiopoietin-1
Pericytes
MicroRNAs
Adventitia
Capillary Permeability
3' Untranslated Regions
Luciferases
Computer Simulation

Keywords

  • angiogenesis
  • microRNA
  • pericytes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Slater, S. C., Jover, E., Martello, A., Mitić, T., Rodriguez-Arabaolaza, I., Vono, R., ... Madeddu, P. (2018). MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1. Molecular Therapy, 26(12), 2823-2837. https://doi.org/10.1016/j.ymthe.2018.08.020

MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1. / Slater, Sadie C.; Jover, Eva; Martello, Andrea; Mitić, Tijana; Rodriguez-Arabaolaza, Iker; Vono, Rosa; Alvino, Valeria V.; Satchell, Simon C.; Spinetti, Gaia; Caporali, Andrea; Madeddu, Paolo.

In: Molecular Therapy, Vol. 26, No. 12, 05.12.2018, p. 2823-2837.

Research output: Contribution to journalArticle

Slater, SC, Jover, E, Martello, A, Mitić, T, Rodriguez-Arabaolaza, I, Vono, R, Alvino, VV, Satchell, SC, Spinetti, G, Caporali, A & Madeddu, P 2018, 'MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1', Molecular Therapy, vol. 26, no. 12, pp. 2823-2837. https://doi.org/10.1016/j.ymthe.2018.08.020
Slater SC, Jover E, Martello A, Mitić T, Rodriguez-Arabaolaza I, Vono R et al. MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1. Molecular Therapy. 2018 Dec 5;26(12):2823-2837. https://doi.org/10.1016/j.ymthe.2018.08.020
Slater, Sadie C. ; Jover, Eva ; Martello, Andrea ; Mitić, Tijana ; Rodriguez-Arabaolaza, Iker ; Vono, Rosa ; Alvino, Valeria V. ; Satchell, Simon C. ; Spinetti, Gaia ; Caporali, Andrea ; Madeddu, Paolo. / MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1. In: Molecular Therapy. 2018 ; Vol. 26, No. 12. pp. 2823-2837.
@article{b680c40a0ebc4e4fbfbfb2d6feb494a4,
title = "MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1",
abstract = "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. Angiopoietin-1 is an important regulator of angiogenesis, yet little is known regarding its transcriptional regulation. Slater et al. (2018) demonstrate angiopoietin-1 induction in pericytes occurs via miR-532-5p regulation of BACH1 and modulation of this pathway promotes changes in endothelial permeability, vascular stability, and angiogenesis.",
keywords = "angiogenesis, microRNA, pericytes",
author = "Slater, {Sadie C.} and Eva Jover and Andrea Martello and Tijana Mitić and Iker Rodriguez-Arabaolaza and Rosa Vono and Alvino, {Valeria V.} and Satchell, {Simon C.} and Gaia Spinetti and Andrea Caporali and Paolo Madeddu",
year = "2018",
month = "12",
day = "5",
doi = "10.1016/j.ymthe.2018.08.020",
language = "English",
volume = "26",
pages = "2823--2837",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Nature Publishing Group",
number = "12",

}

TY - JOUR

T1 - MicroRNA-532-5p Regulates Pericyte Function by Targeting the Transcription Regulator BACH1 and Angiopoietin-1

AU - Slater, Sadie C.

AU - Jover, Eva

AU - Martello, Andrea

AU - Mitić, Tijana

AU - Rodriguez-Arabaolaza, Iker

AU - Vono, Rosa

AU - Alvino, Valeria V.

AU - Satchell, Simon C.

AU - Spinetti, Gaia

AU - Caporali, Andrea

AU - Madeddu, Paolo

PY - 2018/12/5

Y1 - 2018/12/5

N2 - 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. Angiopoietin-1 is an important regulator of angiogenesis, yet little is known regarding its transcriptional regulation. Slater et al. (2018) demonstrate angiopoietin-1 induction in pericytes occurs via miR-532-5p regulation of BACH1 and modulation of this pathway promotes changes in endothelial permeability, vascular stability, and angiogenesis.

AB - 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. Angiopoietin-1 is an important regulator of angiogenesis, yet little is known regarding its transcriptional regulation. Slater et al. (2018) demonstrate angiopoietin-1 induction in pericytes occurs via miR-532-5p regulation of BACH1 and modulation of this pathway promotes changes in endothelial permeability, vascular stability, and angiogenesis.

KW - angiogenesis

KW - microRNA

KW - pericytes

UR - http://www.scopus.com/inward/record.url?scp=85054091166&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85054091166&partnerID=8YFLogxK

U2 - 10.1016/j.ymthe.2018.08.020

DO - 10.1016/j.ymthe.2018.08.020

M3 - Article

C2 - 30274787

AN - SCOPUS:85054091166

VL - 26

SP - 2823

EP - 2837

JO - Molecular Therapy

JF - Molecular Therapy

SN - 1525-0016

IS - 12

ER -