Anti-HB-EGF Antibody-Mediated Delivery of siRNA to Atherosclerotic Lesions in Mice

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Author(s)

    • Tsuchida Shota
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Asakura Masanori
    • Cardiovascular Division, Department of Internal Medicine, Hyogo College of Medicine
    • Hao Hiroyuki
    • Department of Pathology, Nihon University School of Medicine
    • Takashima Seiji
    • Department of Medical Biochemistry, Osaka University Graduate School of Medicine
    • Kitakaze Masafumi
    • Clinical Research and Development, National Cerebral and Cardiovascular Center Research Institute
    • Sakata Yasushi
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Mekada Eisuke
    • Department of Cell Biology, Research Institute for Microbial Diseases, Osaka University
    • Minamino Tetsuo
    • Department of Cardiorenal and Cerebrovascular Medicine, Faculty of Medicine, Kagawa University
    • Matsuzaki Takashi
    • Department of Cardiorenal and Cerebrovascular Medicine, Faculty of Medicine, Kagawa University
    • Yamato Masaki
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Okuda Keiji
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Fu Hai Ying
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Araki Ryo
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Sanada Shoji
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine
    • Asanuma Hiroshi
    • Department of Internal Medicine, Meiji University of Integrative Medicine
    • Asano Yoshihiro
    • Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine

Abstract

<p>For atherosclerotic cardiovascular diseases (ACD), gene therapy may be a potential therapeutic strategy; however, lack of effective and safe methods for gene delivery to atherosclerotic plaques have limited its potential therapeutic applications. To overcome this limitation, we developed a novel antibody-based gene delivery system (anti-HB-EGF/NA vector) by chemically crosslinking antibodies against human heparin-binding epidermal growth factor-like growth factor (HB-EGF). It has been shown to be excessively expressed in human atherosclerotic plaques and NeutrAvidin (NA) for conjugating biotinylated siRNA. Immunofluorescence staining and quantitative flow cytometry analysis using human HB-EGF-expressing cells showed both antibody-mediated selective cellular targeting and efficient intracellular delivery of conjugated biotin-fluorescence. Moreover, we demonstrated antibody-mediated significant and selective gene knockdown via conjugation with anti-HB-EGF/NA vector and biotinylated siRNA (anti-HB-EGF/NA/b-siRNA) <i>in vitro</i>. Furthermore, using high fat-fed human HB-EGF knock-in and apolipoprotein E-knockout (<i>Hbegf</i> hz/hz; <i>Apoe</i>-/-) mice, we demonstrated that the anti-HB-EGF/NA vector, conjugating biotin-fluorescence, increasingly accumulated within the atherosclerotic plaques of the ascending aorta in which human HB-EGF expression levels were highly elevated. Moreover, in response to a single intravenous injection of anti-HB-EGF/NA/b-siRNA in a dose-dependent manner, qPCR analysis of laser-dissected atherosclerotic plaques of the ascending aorta showed significant knockdown of the reporter gene expression. These results suggest that the anti-HB-EGF antibody-mediated siRNA delivery could be a promising delivery system for gene therapy of ACD.</p>

Journal

  • International Heart Journal

    International Heart Journal, 2018

    International Heart Journal Association

Codes

  • NII Article ID (NAID)
    130007505906
  • Text Lang
    ENG
  • ISSN
    1349-2365
  • Data Source
    J-STAGE 
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