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Canadian Journal of Cardiology

Cyclin-Dependent Kinase Inhibitor 2b Controls Fibrosis and Functional Changes in Ischemia-Induced Heart Failure via the BMI1-p15-Rb Signalling Pathway

  • Author Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Wenbo Yang
    Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    Search for articles by this author
  • Author Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Andi Zhang
    Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    Search for articles by this author
  • Author Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Yanxin Han
    Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    Search for articles by this author
  • Xiuxiu Su
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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  • Yanjia Chen
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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  • Weilin Zhao
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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  • Ke Yang
    Correspondence
    Corresponding author: Dr Ke Yang, Department of Cardiology, Ruijin Hospital, and Institute of Cardiovascular Disease, Shanghai Jiao Tong University School of Medicine, No. 197, Ruijin 2nd Road, Shanghai 200025, People’s Republic of China. Tel.: +86-21-64370045, fax: +86-21-64457177.
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Institute of Cardiovascular Disease, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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  • Wei Jin
    Correspondence
    Corresponding author: Dr Wei Jin, Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, No. 197, Ruijin 2nd Road, Shanghai 200025, People’s Republic of China. Tel.: +86-21-64370045, fax: +86-21-64457177.
    Affiliations
    Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    Search for articles by this author
  • Author Footnotes
    ∗ Wenbo Yang, Andi Zhang, and Yanxin Han contributed equally to this work.

      Abstract

      Background

      Cardiac fibrosis is an important cause of heart failure (HF) after myocardial infarction (MI). Cyclin-dependent kinase inhibitor 2b (CDKN2b) regulates the cell cycle by encoding the p15 protein and participates in the development of various tumours. However, the role of CDKN2b/p15 in cardiac fibrosis and HF after MI remains unclear.

      Methods

      Lentivirus was used to induce the silence and overexpression of CDKN2b. Cardiac function was detected with the use of echocardiography. Immunohistochemistry, immunofluorescence, Western blotting, Cell Counting Kit 8, and wound healing assay were used to illustrate the potential mechanism associated with CDKN2b.

      Results

      The p15 protein expression was significantly down-regulated in both human and mouse failing hearts. Cardiac down-regulation of CDKN2b promoted myocardial fibrosis and worsened cardiac function in MI mice, while systemic CDKN2b silencing induced diastolic dysfunction in vivo. In addition, cardiac overexpression of CDKN2b ameliorated cardiac fibrosis and improved cardiac function in MI mice. Mechanistically, silencing CDKN2b gene enhanced the phosphorylation of retinoblastoma (Rb) protein and reinforced the migration and proliferation capabilities of cardiac fibroblasts. B Lymphoma Mo-MLV insertion region 1 homolog (BMI1) was up-regulated in failing heart and inversely regulated the expression of CDKN2b/p15 and the phosphorylation of Rb protein. The BMI1-p15-Rb signalling pathway is a potential mechanism of ischemia-induced cardiac fibrosis and HF.

      Conclusions

      Cardiac fibrosis and heart function could be worsened by the down-regulation and relieved by the up-regulation of CDKN2b/p15 in ischemia-induced HF via regulating the proliferation and migration capabilities of cardiac fibroblasts. These effects could be partially explained by the regulation of the BMI1-p15-Rb signalling pathway.

      Résumé

      Contexte

      La fibrose cardiaque est une importante cause d’insuffisance cardiaque (IC) après la survenue d’un infarctus du myocarde (IM). Le gène CDKN2b (cyclin-dependent kinase inhibitor 2b) participe à la régulation du cycle cellulaire en codant pour la protéine p15 et joue un rôle dans l’apparition de diverses tumeurs. Le rôle de CDKN2b et de la protéine p15 dans la fibrose cardiaque et l’IC après un IM n’est toutefois pas bien connu.

      Méthodologie

      Nous avons utilisé un lentivirus pour induire le silence et la surexpression de CDKN2b. La fonction cardiaque a été évaluée par échocardiographie. Nous avons en outre eu recours à des techniques d’immunohistochimie, d’immunofluorescence et de buvardage de western, ainsi qu’à la trousse CCK-8 (Cell Counting Kit 8) et à une épreuve par scarification pour tenter d’élucider le mode d’action associé à CDKN2b.

      Résultats

      L’expression de la protéine p15 était significativement réduite dans les cœurs humains et murins en insuffisance. La diminution de l’expression de CDKN2b dans le muscle cardiaque a favorisé la fibrose myocardique et détérioré la fonction cardiaque chez des souris ayant subi un IM, tandis que le silence systémique de CDKN2b a induit une dysfonction diastolique in vivo. En outre, la surexpression de CDKN2b dans le muscle cardiaque a réduit la fibrose myocardique et amélioré la fonction cardiaque chez des souris ayant subi un IM. Sur le plan du mode d’action, le silence du gène CDKN2b a rehaussé la phosphorylation de la protéine du rétinoblastome (Rb) et renforcé les capacités de migration et de prolifération des fibroblastes cardiaques. Le gène BMI1 (B Lymphoma Mo-MLV insertion region 1 homolog) était régulé à la hausse dans le tissu cardiaque en insuffisance et a diminué l’expression de CDKN2b/p15 ainsi que la phosphorylation de la protéine du Rb. La voie de signalisation BMI1-p15-Rb pourrait donc jouer un rôle dans la fibrose cardiaque et l’IC d’origine ischémique.

      Conclusions

      La régulation à la baisse de l’expression CDKN2b/p15 pourrait accroître la fibrose cardiaque et affaiblir la fonction cardiaque en cas d’IC d’origine ischémique et, inversement, sa régulation à la hausse pourrait diminuer la fibrose cardiaque et rehausser la fonction cardiaque, ce gène régulant les capacités de prolifération et de migration des fibroblastes cardiaques. Ces effets pourraient s’expliquer en partie par la régulation de la voie de signalisation BMI1-p15-Rb.
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