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Long noncoding TSI attenuates aortic valve calcification by suppressing TGF-β1-induced osteoblastic differentiation of valve interstitial cells

  • Author Footnotes
    1 These authors contributed equally to this work.
    Zongtao Liu
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Author Footnotes
    1 These authors contributed equally to this work.
    Yixuan Wang
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

    Key Laboratory of Organ Transplantation, Ministry of Education, NHC Key Laboratory of Organ Transplantation, Key Laboratory of Organ Transplantation, Chinese Academy of Medical Sciences, Wuhan, China
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.
    Fayuan Liu
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Da Zhu
    Affiliations
    Structural Heart Center, Fuwai Yunnan Cardiovascular Hospital, Kunming Medical University, 528 Shahebei Rd, 65000 Kunming, China
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  • Yuqi Chen
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Wei Yen Yim
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Ke Hu
    Affiliations
    Department of Vascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Zhenqi Rao
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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  • Xiangbin Pan
    Correspondence
    Corresponding authors at: Structural Heart Center, Fuwai Yunnan Cardiovascular Hospital, Kunming Medical University, 528 Shahebei Rd, 65000 Kunming, China
    Affiliations
    Structural Heart Center, Fuwai Yunnan Cardiovascular Hospital, Kunming Medical University, 528 Shahebei Rd, 65000 Kunming, China
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  • Fei Li
    Correspondence
    Corresponding authors at: Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Ave, Wuhan 430022, China.
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

    Key Laboratory of Organ Transplantation, Ministry of Education, NHC Key Laboratory of Organ Transplantation, Key Laboratory of Organ Transplantation, Chinese Academy of Medical Sciences, Wuhan, China
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  • Nianguo Dong
    Correspondence
    Corresponding authors at: Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Ave, Wuhan 430022, China.
    Affiliations
    Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

    Key Laboratory of Organ Transplantation, Ministry of Education, NHC Key Laboratory of Organ Transplantation, Key Laboratory of Organ Transplantation, Chinese Academy of Medical Sciences, Wuhan, China
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.
Published:October 20, 2022DOI:https://doi.org/10.1016/j.metabol.2022.155337

      Highlights

      • Calcific aortic valve disease is an important and common cardiovascular disease.
      • long noncoding TSI expression was upregulated in calcified aortic valves.
      • lncTSI alleviated aortic valve calcification through negative regulation of the TGF-β/Smad3 pathway.
      • These data suggest that long noncoding TSI is a potential therapeutic target.

      Abstract

      Introduction

      Calcific aortic valve disease (CAVD) is an active and cellular-driven fibrocalcific process characterised by differentiation of valve interstitial cells (VICs) towards an osteogenic-like phenotype. A recently identified lncRNA, lncTSI, has been reported to inhibit fibrogenesis through transforming growth factor (TGF)-β/Smad3 pathway. Here, the present study aimed to investigate the role of lncTSI in CAVD.

      Methods

      The effect of TGF-β1 on lncTSI of VICs was measured. TGF-β1, RUNX2 and collagen I expression between calcified aortic valve tissue and normal samples by immunohistochemistry and western blotting. Human VICs were cultured and treated with TGF-β1. SiRNA and pcDNA3.1-lncTSI plasmid transfection were used to silence and overexpress lncTSI in VICs for 48 h, Smads phosphorylation, RUNX2 and collagen I expression were then verified by western blotting. In ApoE−/− mice fed with 0.25 % high-cholesterol diet, AAV2-lncTSI were injected intravenously to observe their effect on the formation of aortic valve calcification.

      Results

      lncTSI was highly expressed in VICs treated with TGF-β1. lncTSI was transcriptionally regulated by Smad3 and reversely inhibited TGF-β1-induced Smad3 phosphorylation and downregulated profibrotic gene expression. Silencing lncTSI increased TGF-β1–induced Smad3 phosphorylation, and subsequently, upregulated RUNX2 and collagen I expressions in VICs. While overexpression of lncTSI reversed the production of RUNX2 and collagen I in VICs. In a mouse CAVD model of 24 week 0.25 % high-cholesterol diet feeding, overexpression of lncTSI significantly reduced calcium deposition, RUNX2, pSmad3, and collagen I expression in aortic valve leaflets, with less aortic valve stenosis.

      Conclusions

      The novel findings of present study suggested that lncTSI alleviated aortic valve calcification through negative regulation of the TGF-β/Smad3 pathway. The results may help elucidate new diagnostic and therapeutic targets to prevent CAVD progression.

      Abbreviations:

      CAVD (calcific aortic valve disease), VICs (valve interstitial cells), TGF-β (transforming growth factor β), lncTSI (long noncoding TSI), IL (interleukin), COL1A1 (type I collagen), RUNX2 (Runt-related Transcription Factor 2), FBS (fetal bovine serum), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), H&E staining (haematoxylin-eosin staining), PBS (phosphate buffer solution), ApoE−/− (apolipoprotein E–deficient), ROS (reactive oxygen species)

      Keywords

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