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Fingerprint of long non-coding RNA regulated by cyclic mechanical stretch in human aortic smooth muscle cells: implications for hypertension

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Abstract

Emerging evidence suggests that long non-coding RNAs (lncRNAs) represent a cellular hub coordinating various cellular processes that are critical in health and disease. Mechanical stress triggers changes in vascular smooth muscle cells (VSMCs) that in turn contribute to pathophysiological changes within the vasculature. We sought to evaluate the role that lncRNAs play in mechanical stretch-induced alterations of human aortic smooth muscle cells (HASMCs). RNA (lncRNA and mRNA) samples isolated from HASMCs that had been subjected to 10 or 20% elongation (1 Hz) for 24 h were profiled with the Arraystar Human LncRNA Microarray V3.0. LncRNA expression was quantified in parallel via qRT-PCR. Of the 30,586 human lncRNAs screened, 580 were differentially expressed (DE, P < 0.05) in stretched HASMCs. Amongst the 26,109 protein-coding transcripts evaluated, 25 of those DE were associated with 25 of the aforementioned DE lncRNAs (P < 0.05). Subsequent Kyoto Encyclopedia of Genes and Genomes analysis revealed that the DE mRNAs were largely associated with the tumor necrosis factor signaling pathway and inflammation. Gene Ontology analysis indicated that the DE mRNAs were associated with cell differentiation, stress response, and response to external stimuli. We describe the first transcriptome profile of stretch-induced changes in HASMCs and provide novel insights into the regulatory switches that may be fundamental in governing aberrant VSMC remodeling.

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Abbreviations

AVIC:

Aortic valve interstitial cell

CAD:

Coronary artery disease

cDNA:

Complementary DNA

CHRF:

Cardiac hypertrophy-related factor

DE:

Differentially expressed

EC:

Endothelial cell

ECM:

Extracellular matrix

FBN-1:

Fibrillin-1

FBSS:

Fetal bovine serum

GO:

Gene ontology

HASMC:

Human aortic smooth muscle cell

KEGG:

Kyoto Encyclopedia of Genes and Genomes

lncRNA:

Long non-coding RNA

Mhrt:

Myosin heavy chain-associated RNA transcripts

ncRNA:

Non-coding RNA

PAD:

Peripheral artery disease

qRT-PCR:

Quantitative real-time PCR

RA:

Rheumatoid arthritis

TNF:

Tumor necrosis factor

VSMC:

Vascular smooth muscle cell

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Acknowledgements

This work was supported by in part by a grant from the Heart and Stroke Foundation of Canada to S. Verma. L. E. Mantella was the recipient of a Canada Graduate Scholarships-Master’s (CGS M) Award and a University of Toronto Fellowship. S. Verma is the Canada Research Chair in Atherosclerosis at the University of Toronto.

Author contributions

LEM, KKS, MA-O, RPJ, and SV designed the studies and the experiments. LEM, PS, CK, and AR conducted the experiments. LEM, AQ, and SM drafted the manuscript. LEM, KKS, NK, JEF, and SV analyzed and interpreted the data. All authors critically edited the manuscript, read and approved the final manuscript.

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Authors and Affiliations

  1. Division of Cardiac Surgery, Keenan Research Centre for Biomedical Science of St. Michael’s Hospital, Toronto, ON, Canada

    Laura-Eve Mantella, Krishna K. Singh, Paul Sandhu, Azza Ramadan, Adrian Quan & Subodh Verma

  2. Division of Vascular Surgery, Keenan Research Centre for Biomedical Science of St. Michael’s Hospital, Toronto, ON, Canada

    Krishna K. Singh & Mohammed Al-Omran

  3. Department of Pharmacology & Toxicology, University of Toronto, Toronto, ON, Canada

    Laura-Eve Mantella & Subodh Verma

  4. Department of Surgery, University of Toronto, Toronto, ON, Canada

    Krishna K. Singh, Mohammed Al-Omran & Subodh Verma

  5. Institute of Medical Science, University of Toronto, Toronto, ON, Canada

    Azza Ramadan & Subodh Verma

  6. Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada

    Nadiya Khyzha & Jason E. Fish

  7. Department of Paediatrics, University of Toronto, Toronto, ON, Canada

    Robert P. Jankov

  8. Department of Physiology, University of Toronto, Toronto, ON, Canada

    Robert P. Jankov

  9. Lung Biology Programme, Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, Toronto, ON, Canada

    Crystal Kantores & Robert P. Jankov

  10. Toronto General Research Institute, University Health Network, Toronto, ON, Canada

    Nadiya Khyzha & Jason E. Fish

  11. Heart & Stroke Richard Lewar Centre of Excellence in Cardiovascular Research, University of Toronto, Toronto, ON, Canada

    Nadiya Khyzha, Jason E. Fish, Robert P. Jankov & Subodh Verma

  12. Department of Surgery, King Saud University, Riyadh, Kingdom of Saudi Arabia

    Mohammed Al-Omran

  13. The King Saud University-Li Ka Shing Collaborative Research Program, Riyadh, Kingdom of Saudi Arabia

    Mohammed Al-Omran

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  1. Laura-Eve Mantella
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Correspondence to Subodh Verma.

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Mantella, LE., Singh, K.K., Sandhu, P. et al. Fingerprint of long non-coding RNA regulated by cyclic mechanical stretch in human aortic smooth muscle cells: implications for hypertension. Mol Cell Biochem 435, 163–173 (2017). https://doi.org/10.1007/s11010-017-3065-2

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