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

• Published in: International Heart Journal Vol. 59; no. 6; pp. 1425 - 1431
• Main Authors: Tsuchida, Shota, Matsuzaki, Takashi, Yamato, Masaki, Okuda, Keiji, Fu, Hai Ying, Araki, Ryo, Sanada, Shoji, Asanuma, Hiroshi, Asano, Yoshihiro, Asakura, Masanori, Hao, Hiroyuki, Takashima, Seiji, Kitakaze, Masafumi, Sakata, Yasushi, Mekada, Eisuke, Minamino, Tetsuo
• Format: Journal Article
• Language: English
• Published: Japan International Heart Journal Association 30-11-2018; Japan Science and Technology Agency
• Subjects: Animals; Antibodies - therapeutic use; Aorta; Apolipoprotein E; Arteriosclerosis; Atherosclerosis; Atherosclerosis - metabolism; Atherosclerosis - therapy; Avidin - immunology; Biotin; Cardiovascular diseases; Coronary vessels; Epidermal growth factor; Flow cytometry; Gene expression; Gene therapy; Gene transfer; Genetic Therapy - methods; Heparin; Heparin-binding EGF-like Growth Factor - immunology; Heparin-binding epidermal growth factor-like growth factor; Humans; Immunofluorescence; Intravenous administration; Mice; Mice, Knockout; Plaques; Reporter gene; RNA, Small Interfering - therapeutic use; siRNA; siRNA delivery; Therapeutic applications; Treatment Outcome; siRNA delivery; Atherosclerosis
• ISSN: 1349-2365; 1349-3299
• DOI: 10.1536/ihj.17-644

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) in vitro. Furthermore, using high fat-fed human HB-EGF knock-in and apolipoprotein E-knockout (Hbegf hz/hz; Apoe-/-) 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.