Mitochondrial regulation of exosomal transported miRNA play a central role in vascular smooth muscle cell proliferation and migration associated with atherogenesis and re-stenosis

Zuhair Al Sulti, Paul Coats

Research output: Contribution to journalMeeting abstract

Abstract

Vascular smooth muscle (VSM) cell proliferation and migration are the hallmark of atherosclerosis and re-stenosis. In this study, we have investigated the role of mitochondrial bioactivity in determining the content of exosomes released by vascular smooth muscle cells and the biological effect these exosomes have on vascular smooth muscle (VSM) cell proliferation and migration.
VSM cells were isolated from 12 week old male Sprague-Dawley rats. Experiments were undertaken using day zero isolated (contractile phenotype), 21 day cultured (synthetic phenotype) and 21 day mitochondrial incompetent (synthetic phenotype- Rho cell). The effect of balloon angioplasty on rat aorta structural remodelling was also studied.
Inhibition of mitochondrial network formation with the DRP1 inhibitor MDivi (10 uM) inhibited angioplasty-dependent remodelling. This observation was confirmed in VSM cell cultures where MDivi significantly reduced proliferation and migration. Total exosomal release was significantly greater in the 21 day cultured cells when compared with quiesced non-proliferating cells and 21 day cultured Rho cells (480±20 ng, 520±10 ng and 420±10 ng per ml respectively) and total exosomal RNA yield was 70.2±10.2 ng/ul, 118.7±2.4 ng/ul and 70.8±4.7 ng/ul respectively.
Mitochondrial function significantly influenced miRNA and mRNA measured in exosomes. When compared with the hyperproliferative synthetic VSM cell miR-21 expression was reduced by 88±12.1% and miR-145 expression increased by 73±19.8%. We also measured a 7-fold decrease and 6.6-fold decrease mTOR, PI3K and 4EBP1 respectively. A significant increase expression of P53, cdkn2a and ROS scavenging proteins including SOD1 and SOD2 were measured in 21 day cultured Rho cells vs. 21 day hyperproliferative VSM cells.
In this study we have further correlated VSM cell hyperproliferative phenotype with mitochondrial function. Moreover, further demonstrated mitochondrial function/VSM cell phenotype with exosomal release and cargo that potentially drives the hyperproliferative/migratory phenotype central to atherosclerosis and re-stenosis.
LanguageEnglish
PagesA1-A1
Number of pages1
JournalHeart
Volume103
Issue numberSuppl 2
DOIs
Publication statusPublished - 30 Apr 2017
Event20th Scottish Cardiovascular Forum - University of Glasgow, Glasgow, United Kingdom
Duration: 4 Feb 20174 Feb 2017

Fingerprint

MicroRNAs
Vascular Smooth Muscle
Smooth Muscle Myocytes
Cell Movement
Atherosclerosis
Pathologic Constriction
Cell Proliferation
Exosomes
Phenotype
Cultured Cells
Balloon Angioplasty
Phosphatidylinositol 3-Kinases
Angioplasty
Sprague Dawley Rats
Aorta
Cell Culture Techniques
RNA
Messenger RNA

Keywords

  • vascular smooth muscle cells
  • mitochondrial bioactivity

Cite this

@article{796a4fbd8e9b4474a5f4afd1d6047a08,
title = "Mitochondrial regulation of exosomal transported miRNA play a central role in vascular smooth muscle cell proliferation and migration associated with atherogenesis and re-stenosis",
abstract = "Vascular smooth muscle (VSM) cell proliferation and migration are the hallmark of atherosclerosis and re-stenosis. In this study, we have investigated the role of mitochondrial bioactivity in determining the content of exosomes released by vascular smooth muscle cells and the biological effect these exosomes have on vascular smooth muscle (VSM) cell proliferation and migration.VSM cells were isolated from 12 week old male Sprague-Dawley rats. Experiments were undertaken using day zero isolated (contractile phenotype), 21 day cultured (synthetic phenotype) and 21 day mitochondrial incompetent (synthetic phenotype- Rho cell). The effect of balloon angioplasty on rat aorta structural remodelling was also studied.Inhibition of mitochondrial network formation with the DRP1 inhibitor MDivi (10 uM) inhibited angioplasty-dependent remodelling. This observation was confirmed in VSM cell cultures where MDivi significantly reduced proliferation and migration. Total exosomal release was significantly greater in the 21 day cultured cells when compared with quiesced non-proliferating cells and 21 day cultured Rho cells (480±20 ng, 520±10 ng and 420±10 ng per ml respectively) and total exosomal RNA yield was 70.2±10.2 ng/ul, 118.7±2.4 ng/ul and 70.8±4.7 ng/ul respectively.Mitochondrial function significantly influenced miRNA and mRNA measured in exosomes. When compared with the hyperproliferative synthetic VSM cell miR-21 expression was reduced by 88±12.1{\%} and miR-145 expression increased by 73±19.8{\%}. We also measured a 7-fold decrease and 6.6-fold decrease mTOR, PI3K and 4EBP1 respectively. A significant increase expression of P53, cdkn2a and ROS scavenging proteins including SOD1 and SOD2 were measured in 21 day cultured Rho cells vs. 21 day hyperproliferative VSM cells.In this study we have further correlated VSM cell hyperproliferative phenotype with mitochondrial function. Moreover, further demonstrated mitochondrial function/VSM cell phenotype with exosomal release and cargo that potentially drives the hyperproliferative/migratory phenotype central to atherosclerosis and re-stenosis.",
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T1 - Mitochondrial regulation of exosomal transported miRNA play a central role in vascular smooth muscle cell proliferation and migration associated with atherogenesis and re-stenosis

AU - Al Sulti, Zuhair

AU - Coats, Paul

PY - 2017/4/30

Y1 - 2017/4/30

N2 - Vascular smooth muscle (VSM) cell proliferation and migration are the hallmark of atherosclerosis and re-stenosis. In this study, we have investigated the role of mitochondrial bioactivity in determining the content of exosomes released by vascular smooth muscle cells and the biological effect these exosomes have on vascular smooth muscle (VSM) cell proliferation and migration.VSM cells were isolated from 12 week old male Sprague-Dawley rats. Experiments were undertaken using day zero isolated (contractile phenotype), 21 day cultured (synthetic phenotype) and 21 day mitochondrial incompetent (synthetic phenotype- Rho cell). The effect of balloon angioplasty on rat aorta structural remodelling was also studied.Inhibition of mitochondrial network formation with the DRP1 inhibitor MDivi (10 uM) inhibited angioplasty-dependent remodelling. This observation was confirmed in VSM cell cultures where MDivi significantly reduced proliferation and migration. Total exosomal release was significantly greater in the 21 day cultured cells when compared with quiesced non-proliferating cells and 21 day cultured Rho cells (480±20 ng, 520±10 ng and 420±10 ng per ml respectively) and total exosomal RNA yield was 70.2±10.2 ng/ul, 118.7±2.4 ng/ul and 70.8±4.7 ng/ul respectively.Mitochondrial function significantly influenced miRNA and mRNA measured in exosomes. When compared with the hyperproliferative synthetic VSM cell miR-21 expression was reduced by 88±12.1% and miR-145 expression increased by 73±19.8%. We also measured a 7-fold decrease and 6.6-fold decrease mTOR, PI3K and 4EBP1 respectively. A significant increase expression of P53, cdkn2a and ROS scavenging proteins including SOD1 and SOD2 were measured in 21 day cultured Rho cells vs. 21 day hyperproliferative VSM cells.In this study we have further correlated VSM cell hyperproliferative phenotype with mitochondrial function. Moreover, further demonstrated mitochondrial function/VSM cell phenotype with exosomal release and cargo that potentially drives the hyperproliferative/migratory phenotype central to atherosclerosis and re-stenosis.

AB - Vascular smooth muscle (VSM) cell proliferation and migration are the hallmark of atherosclerosis and re-stenosis. In this study, we have investigated the role of mitochondrial bioactivity in determining the content of exosomes released by vascular smooth muscle cells and the biological effect these exosomes have on vascular smooth muscle (VSM) cell proliferation and migration.VSM cells were isolated from 12 week old male Sprague-Dawley rats. Experiments were undertaken using day zero isolated (contractile phenotype), 21 day cultured (synthetic phenotype) and 21 day mitochondrial incompetent (synthetic phenotype- Rho cell). The effect of balloon angioplasty on rat aorta structural remodelling was also studied.Inhibition of mitochondrial network formation with the DRP1 inhibitor MDivi (10 uM) inhibited angioplasty-dependent remodelling. This observation was confirmed in VSM cell cultures where MDivi significantly reduced proliferation and migration. Total exosomal release was significantly greater in the 21 day cultured cells when compared with quiesced non-proliferating cells and 21 day cultured Rho cells (480±20 ng, 520±10 ng and 420±10 ng per ml respectively) and total exosomal RNA yield was 70.2±10.2 ng/ul, 118.7±2.4 ng/ul and 70.8±4.7 ng/ul respectively.Mitochondrial function significantly influenced miRNA and mRNA measured in exosomes. When compared with the hyperproliferative synthetic VSM cell miR-21 expression was reduced by 88±12.1% and miR-145 expression increased by 73±19.8%. We also measured a 7-fold decrease and 6.6-fold decrease mTOR, PI3K and 4EBP1 respectively. A significant increase expression of P53, cdkn2a and ROS scavenging proteins including SOD1 and SOD2 were measured in 21 day cultured Rho cells vs. 21 day hyperproliferative VSM cells.In this study we have further correlated VSM cell hyperproliferative phenotype with mitochondrial function. Moreover, further demonstrated mitochondrial function/VSM cell phenotype with exosomal release and cargo that potentially drives the hyperproliferative/migratory phenotype central to atherosclerosis and re-stenosis.

KW - vascular smooth muscle cells

KW - mitochondrial bioactivity

UR - http://heart.bmj.com/content/103/Suppl_2/A1.1

U2 - 10.1136/heartjnl-2017-311433.1

DO - 10.1136/heartjnl-2017-311433.1

M3 - Meeting abstract

VL - 103

SP - A1-A1

JO - Heart

T2 - Heart

JF - Heart

SN - 1355-6037

IS - Suppl 2

ER -