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Dicer generates a regulatory microRNA network in smooth muscle cells that limits neointima formation during vascular repair

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Abstract

MicroRNAs (miRNAs) coordinate vascular repair by regulating injury-induced gene expression in vascular smooth muscle cells (SMCs) and promote the transition of SMCs from a contractile to a proliferating phenotype. However, the effect of miRNA expression in SMCs on neointima formation is unclear. Therefore, we studied the role of miRNA biogenesis by Dicer in SMCs in vascular repair. Following wire-induced injury to carotid arteries of Apolipoprotein E knockout (Apoe −/−) mice, miRNA microarray analysis revealed that the most significantly regulated miRNAs, such as miR-222 and miR-21-3p, were upregulated. Conditional deletion of Dicer in SMCs increased neointima formation by reducing SMC proliferation in Apoe −/− mice, and decreased mainly the expression of miRNAs, such as miR-147 and miR-100, which were not upregulated following vascular injury. SMC-specific deletion of Dicer promoted growth factor and inflammatory signaling and regulated a miRNA–target interaction network in injured arteries that was enriched in anti-proliferative miRNAs. The most connected miRNA in this network was miR-27a-3p [e.g., with Rho guanine nucleotide exchange factor 26 (ARHGEF26)], which was expressed in medial and neointimal SMCs in a Dicer-dependent manner. In vitro, miR-27a-3p suppresses ARHGEF26 expression and inhibits SMC proliferation by interacting with a conserved binding site in the 3′ untranslated region of ARHGEF26 mRNA. We propose that Dicer expression in SMCs plays an essential role in vascular repair by generating anti-proliferative miRNAs, such as miR-27a-3p, to prevent vessel stenosis due to exaggerated neointima formation.

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Abbreviations

Apoe :

Apolipoprotein E

ARHGEF26 :

Rho guanine nucleotide exchange factor 26

CHST1 :

Carbohydrate (keratan sulfate Gal-6) sulfotransferase 1

DAPI:

4’,6-Diamidino-2-phenylindole

DLL4 :

Delta-like 4

DMEM:

Dulbecco′s modified eagle medium

EC:

Endothelial cell

EGF:

Epidermal growth factor

HASMC:

Human aortic smooth muscle cell

HEK293:

Human embryonic kidney 293 cell

HFD:

High-fat diet

IGFBP3 :

Insulin-like growth-factor-binding protein 3

IL-1β:

Interleukin-1β

KO:

Knockout

LNA:

Locked nucleic acid

microRNA:

miRNA, miR

MYH11 :

Myosin, heavy chain 11, smooth muscle

NF-κB:

Nuclear factor of kappa light polypeptide gene enhancer in B-cells

OIT3 :

Oncoprotein-induced transcript 3

PDGF:

Platelet-derived growth factor

miRISC:

miRNA-induced-silencing complex

SH3BGRL2 :

SH3-domain-binding glutamate-rich protein-like 2

SMA:

Smooth muscle actin

SMC:

Smooth muscle cell

TAGLN :

Transgelin

TNFα:

Tumor necrosis factor α

TNRC6A:

Argonaute and trinucleotide repeat containing 6A

TSB:

Target site blocker

UTR:

Untranslated region

WT:

Wild type

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Acknowledgments

This work has been funded by the German Research Foundation (DFG) as part of the Collaborative Research Center 1123 (B04) and by the German Center for Cardiovascular Research (MHA VD1.2). The authors declare no competing financial interests.

Author information

Author notes

  1. Zhe Zhou

    Present address: The Genomics Center of AMMS, Beijing Institute of Radiation Medicine, Beijing, 100850, People’s Republic of China

  2. Pallavi Subramanian

    Present address: Department of Clinical Pathobiochemistry, University Clinic Carl Gustav Carus, Dresden University of Technology, 01307, Dresden, Germany

Authors and Affiliations

  1. Institute for Cardiovascular Prevention, Ludwig-Maximilians-University Munich, 80336, Munich, Germany

    Farima Zahedi, Maliheh Nazari-Jahantigh, Pallavi Subramanian, Yuanyuan Wei, Sabine Steffens, Christian Weber & Andreas Schober

  2. DZHK (German Center for Cardiovascular Research), Partner Site Munich Heart Alliance, 80802, Munich, Germany

    Maliheh Nazari-Jahantigh, Yuanyuan Wei, Sabine Steffens, Christian Weber & Andreas Schober

  3. Institute for Molecular Cardiovascular Research, RWTH Aachen University, 52074, Aachen, Germany

    Zhe Zhou & Andreas Schober

  4. European Vascular Center Aachen-Maastricht, Medical University Maastricht, 6229 HX, Maastricht, The Netherlands

    Jochen Grommes

  5. European Vascular Center Aachen-Maastricht, RWTH Aachen University, 52074, Aachen, Germany

    Jochen Grommes

  6. Max Planck Institute for Heart and Lung Research, 61231, Bad Nauheim, Germany

    Stefan Offermanns

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  1. Farima Zahedi
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Corresponding author

Correspondence to Andreas Schober.

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F. Zahedi and M. Nazari-Jahantigh are co-first authors.

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Zahedi, F., Nazari-Jahantigh, M., Zhou, Z. et al. Dicer generates a regulatory microRNA network in smooth muscle cells that limits neointima formation during vascular repair. Cell. Mol. Life Sci. 74, 359–372 (2017). https://doi.org/10.1007/s00018-016-2349-0

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