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New mechanistic insights to PLOD1-mediated human vascular disease

  • Author Footnotes
    # Indicates Co-first authorship
    Sara N Koenig
    Footnotes
    # Indicates Co-first authorship
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Author Footnotes
    # Indicates Co-first authorship
    Omer Cavus
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    # Indicates Co-first authorship
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Jordan Williams
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Matthew Bernier
    Affiliations
    The Ohio State University Mass Spectrometry and Proteomics Facility, Office of Research, Columbus, Ohio
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  • Jeff Tonniges
    Affiliations
    The Ohio State University Microscopy and Imaging Facility (CMIF), Office of Research, Columbus, Ohio
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  • Holly Sucharski
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Trevor Dew
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Muhannad Akel
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Peter Baker
    Affiliations
    Nationwide Children's Hospital, Department of Pathology, Columbus, Ohio
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  • Francesca Madiai
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Francesca De Giorgi
    Affiliations
    The Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, Italy
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  • Luigi Scietti
    Affiliations
    The Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, Italy
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  • Silvia Faravelli
    Affiliations
    The Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, Italy
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  • Federico Forneris
    Affiliations
    The Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, Italy
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  • Peter J Mohler
    Affiliations
    The Ohio State University College of Medicine, Department of Physiology and Cell Biology, Columbus, Ohio

    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio
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  • Elisa A Bradley
    Correspondence
    Reprint requests: Elisa A Bradley, The Ohio State University College of Medicine, 473 W. 12th Avenue DHLRI Suite 200 Columbus, OH 43220.
    Affiliations
    The Dorothy Davis Heart and Lung Research Institute and the Frick Center for Heart Failure and Arrhythmia, The Ohio State University, Columbus, Ohio

    The Ohio State University College of Medicine and Wexner Medical Center, Division of Cardiovascular Medicine, Department of Internal Medicine, Columbus, Ohio
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  • Author Footnotes
    # Indicates Co-first authorship
Published:August 13, 2021DOI:https://doi.org/10.1016/j.trsl.2021.08.002
      Heritable thoracic aortic disease and familial thoracic aortic aneurysm/dissection are important causes of human morbidity/mortality, most without identifiable genetic cause. In a family with familial thoracic aortic aneurysm/dissection, we identified a missense p. (Ser178Arg) variant in PLOD1 segregating with disease, and evaluated PLOD1 enzymatic activity, collagen characteristics and in human aortic vascular smooth muscle cells, studied the effect on function. Comparison with homologous PLOD3 enzyme indicated that the pathogenic variant may affect the N-terminal glycosyltransferase domain, suggesting unprecedented PLOD1 activity. In vitro assays demonstrated that wild-type PLOD1 is capable of processing UDP-glycan donor substrates, and that the variant affects the folding stability of the glycosyltransferase domain and associated enzymatic functions. The PLOD1 substrate lysine was elevated in the proband, however the enzymatic product hydroxylysine and total collagen content was not different, albeit despite collagen fibril narrowing and preservation of collagen turnover. In VSMCs overexpressing wild-type PLOD1, there was upregulation in procollagen gene expression (secretory function) which was attenuated in the variant, consistent with loss-of-function. In comparison, si-PLOD1 cells demonstrated hypercontractility and upregulation of contractile markers, providing evidence for phenotypic switching. Together, the findings suggest that the PLOD1 product is preserved, however newly identified glucosyltransferase activity of PLOD1 appears to be affected by folding stability of the variant, and is associated with compensatory vascular smooth muscle cells phenotypic switching to support collagen production, albeit with less robust fibril girth. Future studies should focus on the impact of PLOD1 folding/variant stability on the tertiary structure of collagen and ECM interactions.

      Abbreviations:

      CSA/Ht (cross-sectional-area to height ratio), ECM (extracellular matrix), ECU (elastin-contractile unit), FTAAD (familial thoracic aortic aneurysm and dissection), GT (glycosyltransferase), HTAD (heritable thoracic aortic disease), kEDS (kyphoscoliotic Ehlers Danlos syndrome), PLOD1 (procollagen-lysine 2-oxoglutarate 5-dioxygenase 1), VSMC ((human aortic) Vascular smooth muscle cell), VUS (variant of unknown significance)
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