The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling

Amira D. Mahmoud, Margaret D. Ballantyne, Vladislav Miscianinov, Karine Pinel, John Hung, Jessica P. Scanlon, Jean Iyinikkel, Jakub Kaczynski, Adriana S. Tavares, Angela C. Bradshaw, Nicholas L. Mills, David E. Newby, Andrea Caporali, Gwyn W. Gould, Sarah J. George, Igor Ulitsky, Judith C. Sluimer, Julie Rodor, Andrew H. Baker

Research output: Contribution to journalArticle

Abstract

RATIONALE:
In response to blood vessel wall injury, aberrant proliferation of vascular smooth muscle cells (SMCs) causes pathological remodeling. However, the controlling mechanisms are not completely understood.
OBJECTIVE:
We recently showed that the human long noncoding RNA, SMILR, promotes vascular SMCs proliferation by a hitherto unknown mechanism. Here, we assess the therapeutic potential of SMILR inhibition and detail the molecular mechanism of action.
METHODS AND RESULTS:
We used deep RNA-sequencing of human saphenous vein SMCs stimulated with IL (interleukin)-1α and PDGF (platelet-derived growth factor)-BB with SMILR knockdown (siRNA) or overexpression (lentivirus), to identify SMILR-regulated genes. This revealed a SMILR-dependent network essential for cell cycle progression. In particular, we found using the fluorescent ubiquitination-based cell cycle indicator viral system that SMILR regulates the late mitotic phase of the cell cycle and cytokinesis with SMILR knockdown resulting in ≈10% increase in binucleated cells. SMILR pulldowns further revealed its potential molecular mechanism, which involves an interaction with the mRNA of the late mitotic protein CENPF (centromere protein F) and the regulatory Staufen1 RNA-binding protein. SMILR and this downstream axis were also found to be activated in the human ex vivo vein graft pathological model and in primary human coronary artery SMCs and atherosclerotic plaques obtained at carotid endarterectomy. Finally, to assess the therapeutic potential of SMILR, we used a novel siRNA approach in the ex vivo vein graft model (within the 30 minutes clinical time frame that would occur between harvest and implant) to assess the reduction of proliferation by EdU incorporation. SMILR knockdown led to a marked decrease in proliferation from ≈29% in controls to ≈5% with SMILR depletion.
CONCLUSIONS:
Collectively, we demonstrate that SMILR is a critical mediator of vascular SMC proliferation via direct regulation of mitotic progression. Our data further reveal a potential SMILR-targeting intervention to limit atherogenesis and adverse vascular remodeling.
LanguageEnglish
Pages535-551
Number of pages17
JournalCirculation Research
Volume125
Issue number5
Early online date24 Jul 2019
DOIs
Publication statusPublished - 16 Aug 2019

Fingerprint

Smooth Muscle Myocytes
Cell Cycle
Messenger RNA
Vascular Smooth Muscle
Small Interfering RNA
Veins
Cell Proliferation
Long Noncoding RNA
RNA Sequence Analysis
Transplants
High-Throughput Nucleotide Sequencing
Lentivirus
Interleukin-1alpha
RNA-Binding Proteins
Cytokinesis
Carotid Endarterectomy
Ubiquitination
Saphenous Vein
Atherosclerotic Plaques
Blood Vessels

Keywords

  • blood vessels
  • cell cycle
  • growth factors
  • interleukins
  • muscle cells
  • noncoding RNA
  • saphenous vein

Cite this

Mahmoud, Amira D. ; Ballantyne, Margaret D. ; Miscianinov, Vladislav ; Pinel, Karine ; Hung, John ; Scanlon, Jessica P. ; Iyinikkel, Jean ; Kaczynski, Jakub ; Tavares, Adriana S. ; Bradshaw, Angela C. ; Mills, Nicholas L. ; Newby, David E. ; Caporali, Andrea ; Gould, Gwyn W. ; George, Sarah J. ; Ulitsky, Igor ; Sluimer, Judith C. ; Rodor, Julie ; Baker, Andrew H. / The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling. In: Circulation Research. 2019 ; Vol. 125, No. 5. pp. 535-551.
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title = "The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling",
abstract = "RATIONALE:In response to blood vessel wall injury, aberrant proliferation of vascular smooth muscle cells (SMCs) causes pathological remodeling. However, the controlling mechanisms are not completely understood.OBJECTIVE:We recently showed that the human long noncoding RNA, SMILR, promotes vascular SMCs proliferation by a hitherto unknown mechanism. Here, we assess the therapeutic potential of SMILR inhibition and detail the molecular mechanism of action.METHODS AND RESULTS:We used deep RNA-sequencing of human saphenous vein SMCs stimulated with IL (interleukin)-1α and PDGF (platelet-derived growth factor)-BB with SMILR knockdown (siRNA) or overexpression (lentivirus), to identify SMILR-regulated genes. This revealed a SMILR-dependent network essential for cell cycle progression. In particular, we found using the fluorescent ubiquitination-based cell cycle indicator viral system that SMILR regulates the late mitotic phase of the cell cycle and cytokinesis with SMILR knockdown resulting in ≈10{\%} increase in binucleated cells. SMILR pulldowns further revealed its potential molecular mechanism, which involves an interaction with the mRNA of the late mitotic protein CENPF (centromere protein F) and the regulatory Staufen1 RNA-binding protein. SMILR and this downstream axis were also found to be activated in the human ex vivo vein graft pathological model and in primary human coronary artery SMCs and atherosclerotic plaques obtained at carotid endarterectomy. Finally, to assess the therapeutic potential of SMILR, we used a novel siRNA approach in the ex vivo vein graft model (within the 30 minutes clinical time frame that would occur between harvest and implant) to assess the reduction of proliferation by EdU incorporation. SMILR knockdown led to a marked decrease in proliferation from ≈29{\%} in controls to ≈5{\%} with SMILR depletion.CONCLUSIONS:Collectively, we demonstrate that SMILR is a critical mediator of vascular SMC proliferation via direct regulation of mitotic progression. Our data further reveal a potential SMILR-targeting intervention to limit atherogenesis and adverse vascular remodeling.",
keywords = "blood vessels, cell cycle, growth factors, interleukins, muscle cells, noncoding RNA, saphenous vein",
author = "Mahmoud, {Amira D.} and Ballantyne, {Margaret D.} and Vladislav Miscianinov and Karine Pinel and John Hung and Scanlon, {Jessica P.} and Jean Iyinikkel and Jakub Kaczynski and Tavares, {Adriana S.} and Bradshaw, {Angela C.} and Mills, {Nicholas L.} and Newby, {David E.} and Andrea Caporali and Gould, {Gwyn W.} and George, {Sarah J.} and Igor Ulitsky and Sluimer, {Judith C.} and Julie Rodor and Baker, {Andrew H.}",
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doi = "10.1161/CIRCRESAHA.119.314876",
language = "English",
volume = "125",
pages = "535--551",
journal = "Circulation Research",
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Mahmoud, AD, Ballantyne, MD, Miscianinov, V, Pinel, K, Hung, J, Scanlon, JP, Iyinikkel, J, Kaczynski, J, Tavares, AS, Bradshaw, AC, Mills, NL, Newby, DE, Caporali, A, Gould, GW, George, SJ, Ulitsky, I, Sluimer, JC, Rodor, J & Baker, AH 2019, 'The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling' Circulation Research, vol. 125, no. 5, pp. 535-551. https://doi.org/10.1161/CIRCRESAHA.119.314876

The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling. / Mahmoud, Amira D.; Ballantyne, Margaret D.; Miscianinov, Vladislav; Pinel, Karine; Hung, John; Scanlon, Jessica P.; Iyinikkel, Jean; Kaczynski, Jakub; Tavares, Adriana S.; Bradshaw, Angela C.; Mills, Nicholas L.; Newby, David E.; Caporali, Andrea; Gould, Gwyn W.; George, Sarah J.; Ulitsky, Igor; Sluimer, Judith C.; Rodor, Julie; Baker, Andrew H.

In: Circulation Research, Vol. 125, No. 5, 16.08.2019, p. 535-551.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The human-specific and smooth muscle cell-enriched LncRNA SMILR promotes proliferation by regulating mitotic CENPF mRNA and drives cell-cycle progression which can be targeted to limit vascular remodeling

AU - Mahmoud, Amira D.

AU - Ballantyne, Margaret D.

AU - Miscianinov, Vladislav

AU - Pinel, Karine

AU - Hung, John

AU - Scanlon, Jessica P.

AU - Iyinikkel, Jean

AU - Kaczynski, Jakub

AU - Tavares, Adriana S.

AU - Bradshaw, Angela C.

AU - Mills, Nicholas L.

AU - Newby, David E.

AU - Caporali, Andrea

AU - Gould, Gwyn W.

AU - George, Sarah J.

AU - Ulitsky, Igor

AU - Sluimer, Judith C.

AU - Rodor, Julie

AU - Baker, Andrew H.

PY - 2019/8/16

Y1 - 2019/8/16

N2 - RATIONALE:In response to blood vessel wall injury, aberrant proliferation of vascular smooth muscle cells (SMCs) causes pathological remodeling. However, the controlling mechanisms are not completely understood.OBJECTIVE:We recently showed that the human long noncoding RNA, SMILR, promotes vascular SMCs proliferation by a hitherto unknown mechanism. Here, we assess the therapeutic potential of SMILR inhibition and detail the molecular mechanism of action.METHODS AND RESULTS:We used deep RNA-sequencing of human saphenous vein SMCs stimulated with IL (interleukin)-1α and PDGF (platelet-derived growth factor)-BB with SMILR knockdown (siRNA) or overexpression (lentivirus), to identify SMILR-regulated genes. This revealed a SMILR-dependent network essential for cell cycle progression. In particular, we found using the fluorescent ubiquitination-based cell cycle indicator viral system that SMILR regulates the late mitotic phase of the cell cycle and cytokinesis with SMILR knockdown resulting in ≈10% increase in binucleated cells. SMILR pulldowns further revealed its potential molecular mechanism, which involves an interaction with the mRNA of the late mitotic protein CENPF (centromere protein F) and the regulatory Staufen1 RNA-binding protein. SMILR and this downstream axis were also found to be activated in the human ex vivo vein graft pathological model and in primary human coronary artery SMCs and atherosclerotic plaques obtained at carotid endarterectomy. Finally, to assess the therapeutic potential of SMILR, we used a novel siRNA approach in the ex vivo vein graft model (within the 30 minutes clinical time frame that would occur between harvest and implant) to assess the reduction of proliferation by EdU incorporation. SMILR knockdown led to a marked decrease in proliferation from ≈29% in controls to ≈5% with SMILR depletion.CONCLUSIONS:Collectively, we demonstrate that SMILR is a critical mediator of vascular SMC proliferation via direct regulation of mitotic progression. Our data further reveal a potential SMILR-targeting intervention to limit atherogenesis and adverse vascular remodeling.

AB - RATIONALE:In response to blood vessel wall injury, aberrant proliferation of vascular smooth muscle cells (SMCs) causes pathological remodeling. However, the controlling mechanisms are not completely understood.OBJECTIVE:We recently showed that the human long noncoding RNA, SMILR, promotes vascular SMCs proliferation by a hitherto unknown mechanism. Here, we assess the therapeutic potential of SMILR inhibition and detail the molecular mechanism of action.METHODS AND RESULTS:We used deep RNA-sequencing of human saphenous vein SMCs stimulated with IL (interleukin)-1α and PDGF (platelet-derived growth factor)-BB with SMILR knockdown (siRNA) or overexpression (lentivirus), to identify SMILR-regulated genes. This revealed a SMILR-dependent network essential for cell cycle progression. In particular, we found using the fluorescent ubiquitination-based cell cycle indicator viral system that SMILR regulates the late mitotic phase of the cell cycle and cytokinesis with SMILR knockdown resulting in ≈10% increase in binucleated cells. SMILR pulldowns further revealed its potential molecular mechanism, which involves an interaction with the mRNA of the late mitotic protein CENPF (centromere protein F) and the regulatory Staufen1 RNA-binding protein. SMILR and this downstream axis were also found to be activated in the human ex vivo vein graft pathological model and in primary human coronary artery SMCs and atherosclerotic plaques obtained at carotid endarterectomy. Finally, to assess the therapeutic potential of SMILR, we used a novel siRNA approach in the ex vivo vein graft model (within the 30 minutes clinical time frame that would occur between harvest and implant) to assess the reduction of proliferation by EdU incorporation. SMILR knockdown led to a marked decrease in proliferation from ≈29% in controls to ≈5% with SMILR depletion.CONCLUSIONS:Collectively, we demonstrate that SMILR is a critical mediator of vascular SMC proliferation via direct regulation of mitotic progression. Our data further reveal a potential SMILR-targeting intervention to limit atherogenesis and adverse vascular remodeling.

KW - blood vessels

KW - cell cycle

KW - growth factors

KW - interleukins

KW - muscle cells

KW - noncoding RNA

KW - saphenous vein

U2 - 10.1161/CIRCRESAHA.119.314876

DO - 10.1161/CIRCRESAHA.119.314876

M3 - Article

VL - 125

SP - 535

EP - 551

JO - Circulation Research

T2 - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 5

ER -