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Free DNA precipitates calcium phosphate apatite crystals in the arterial wall in vivo

  • Raphaël Coscas
    Correspondence
    Corresponding author. Department of Vascular Surgery, Ambroise Paré University Hospital, AP-HP 9, Avenue Charles de Gaulle, 92104 Boulogne-Billancourt, France.
    Affiliations
    UMR 1148, Inserm-Paris7, Denis Diderot University, Xavier Bichat Hospital, 75018 Paris, France

    UMR 1173, Inserm-Paris11, Faculty of Health Sciences Simone Veil, Versailles Saint-Quentin-en-Yvelines University, Paris-Saclay University, 78180 Montigny-le-Bretonneux, France

    Department of Vascular Surgery, Ambroise Paré University Hospital, AP-HP, 92104 Boulogne-Billancourt, France

    UMR 1018, Inserm-Paris11, CESP, Versailles Saint-Quentin-en-Yvelines University, Paris-Saclay University, Paul Brousse Hospital, 94807 Villejuif, France
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  • Marie Bensussan
    Affiliations
    UMR 1148, Inserm-Paris7, Denis Diderot University, Xavier Bichat Hospital, 75018 Paris, France

    Jules Verne University of Picardie and Department of Vascular Surgery, Amiens-Picardie University Hospital, 80054 Amiens, France
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  • Marie-Paule Jacob
    Affiliations
    UMR 1148, Inserm-Paris7, Denis Diderot University, Xavier Bichat Hospital, 75018 Paris, France
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  • Liliane Louedec
    Affiliations
    UMR 1148, Inserm-Paris7, Denis Diderot University, Xavier Bichat Hospital, 75018 Paris, France
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  • Ziad Massy
    Affiliations
    UMR 1018, Inserm-Paris11, CESP, Versailles Saint-Quentin-en-Yvelines University, Paris-Saclay University, Paul Brousse Hospital, 94807 Villejuif, France

    Department of Nephrology, Ambroise Paré University Hospital, AP-HP, 92104 Boulogne-Billancourt, France
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  • Jeremy Sadoine
    Affiliations
    Laboratoire EA2496 et Plateforme d’Imagerie du Vivant (PIV), Paris Descartes University, Sorbonne Paris Cité, Montrouge, France
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  • Michel Daudon
    Affiliations
    Service d'Explorations Fonctionnelles Multidisciplinaires, AP-HP, Hôpital Tenon, Paris, France

    Inserm, UMR S 1155, Hôpital Tenon, Paris, France
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  • Catherine Chaussain
    Affiliations
    Laboratoire EA2496 et Plateforme d’Imagerie du Vivant (PIV), Paris Descartes University, Sorbonne Paris Cité, Montrouge, France
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  • Dominique Bazin
    Affiliations
    CNRS, Laboratoire de Chimie de la Matière Condensée de Paris, UPMC, Collège de France, Paris, France

    Laboratoire de Physique des Solides, Université Paris XI, Orsay, France
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  • Jean-Baptiste Michel
    Affiliations
    UMR 1148, Inserm-Paris7, Denis Diderot University, Xavier Bichat Hospital, 75018 Paris, France
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      Highlights

      • Free DNA colocalizes with calcifications in human and rat arteries.
      • EDTA calcium chelation allowed to identify free DNA as background of calcification.
      • A calcification model based on intra-aortic free DNA infusion is presented.
      • Free DNA may serve as an anionic platform precipitating calcium phosphate.

      Abstract

      Background and aims

      The arterial wall calcium score and circulating free DNA levels are now used in clinical practice as biomarkers of cardiovascular risk. Calcium phosphate apatite retention in the arterial wall necessitates precipitation on an anionic platform. Here, we explore the role of tissue-free DNA as such a platform.

      Methods

      The first step consisted of histological observation of samples from human and rat calcified arteries. Various stains were used to evaluate colocalization of free DNA with calcified tissue (alizarin red, fluorescent Hoechst, DNA immunostaining and TUNEL assay). Sections were treated by EDTA to reveal calcification background. Secondly, a rat model of vascular calcifications induced by intra-aortic infusions of free DNA and elastase + free DNA was developed. Rat aortas underwent a micro-CT for calcium score calculation at 3 weeks. Rat and human calcifications were qualitatively characterized using μFourier Transform Infrared Spectroscopy (μFTIR) and Field Emission-Scanning Electron Microscopy (FE-SEM).

      Results

      Our histological study shows colocalization of calcified arterial plaques with free DNA. In the intra-aortic infusion model, free DNA was able to penetrate into the arterial wall and induce calcifications whereas no microscopic calcification was seen in control aortas. The calcification score in the elastase + free DNA group was significantly higher than in the control groups. Qualitative evaluation with μFTIR and FE-SEM demonstrated typical calcium phosphate retention in human and rat arterial specimens.

      Conclusions

      This translational study demonstrates that free DNA could be involved in arterial calcification formation by precipitating calcium phosphate apatite crystals in the vessel wall.

      Graphical abstract

      Keywords

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