Sandra Pinkert1,2, Markian Pryshliak1, Kathleen Pappritz3,4, Klaus Knoch5, Ahmet Hazini1, Babette Dieringer1, Katrin Schaar1, Fengquan Dong4, Luisa Hinze1, Jie Lin4, Dirk Lassner6, Robert Klopfleisch7, Michele Solimena5, Carsten Tschöpe8, Ziya Kaya9, Muhammad El-Shafeey3,10, Antje Beling2, Jens Kurreck1, Sophie Van Linthout3,4,8, Karin Klingel11, Henry Fechner1. 1. Department of Applied Biochemistry, Institute of Biotechnology, Technische Universität Berlin, Gustav-Meyer-Allee 25, 15533 Berlin, Germany. 2. Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Institute of Biochemistry, Virchowweg 6, 10117 Berlin, Germany. 3. Berlin-Brandenburger Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow Klinikum (CVK), Föhrer Str. 15, 13353 Berlin, Germany. 4. German Center for Cardiovascular Research (DZHK), Partner Site Berlin-Charité, Oudenarder Straße 16, 13316 Berlin, Germany. 5. Faculty of Medicine, Paul Langerhans Institute Dresden of the Helmholtz Center Munich at University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, 01307 Dresden, Germany. 6. Institut Kardiale Diagnostik und Therapie (IKDT), Moltkestraße 31, 12203 Berlin, Germany. 7. Institute of Veterinary Pathology, Freie Universität Berlin, Kaiserswerther Str. 16-18, 14195 Berlin, Germany. 8. Department of Cardiology, Charité-Universitätsmedizin Berlin, Campus Virchow Klinikum (CVK), Charitéplatz 1, 10117 Berlin, Germany. 9. Department of Medicine III, University of Heidelberg, 69120 Heidelberg, Germany; DZHK (German Centre for Cardiovascular Research), Partner Site Heidelberg/Mannheim, University of Heidelberg, 69120 Heidelberg, Germany. 10. Medical Biotechnology Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications, Alexandria, Egypt. 11. Cardiopathology, Institute for Pathology and Neuropathology, University Hospital Tuebingen, Liebermeisterstr. 8, 72076 Tübingen, Germany.
Abstract
AIMS: The coxsackievirus B3 (CVB3) mouse myocarditis model is the standard model for investigation of virus-induced myocarditis but the pancreas, rather than the heart, is the most susceptible organ in mouse. The aim of this study was to develop a CVB3 mouse myocarditis model in which animals develop myocarditis while attenuating viral infection of the pancreas and the development of severe pancreatitis. METHODS AND RESULTS: We developed the recombinant CVB3 variant H3N-375TS by inserting target sites (TS) of miR-375, which is specifically expressed in the pancreas, into the 3'UTR of the genome of the pancreo- and cardiotropic CVB3 variant H3. In vitro evaluation showed that H3N-375TS was suppressed in pancreatic miR-375-expressing EndoC-βH1 cells >5 log10, whereas its replication was not suppressed in isolated primary embryonic mouse cardiomyocytes. In vivo, intraperitoneal (i.p.) administration of H3N-375TS to NMRI mice did not result in pancreatic or cardiac infection. In contrast, intravenous (i.v.) administration of H3N-375TS to NMRI and Balb/C mice resulted in myocardial infection and acute and chronic myocarditis, whereas the virus was not detected in the pancreas and the pancreatic tissue was not damaged. Acute myocarditis was characterized by myocardial injury, inflammation with mononuclear cells, induction of proinflammatory cytokines, and detection of replicating H3N-375TS in the heart. Mice with chronic myocarditis showed myocardial fibrosis and persistence of H3N-375TS genomic RNA but no replicating virus in the heart. Moreover, H3N-375TS infected mice showed distinctly less suffering compared with mice that developed pancreatitis and myocarditis after i.p. or i.v application of control virus. CONCLUSION: In this study, we demonstrate that by use of the miR-375-sensitive CVB3 variant H3N-375TS, CVB3 myocarditis can be established without the animals developing severe systemic infection and pancreatitis. As the H3N-375TS myocarditis model depends on pancreas-attenuated H3N-375TS, it can easily be used in different mouse strains and for various applications. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: The coxsackievirus B3 (CVB3) mousemyocarditis model is the standard model for investigation of virus-induced myocarditis but the pancreas, rather than the heart, is the most susceptible organ in mouse. The aim of this study was to develop a CVB3mousemyocarditis model in which animals develop myocarditis while attenuating viral infection of the pancreas and the development of severe pancreatitis. METHODS AND RESULTS: We developed the recombinant CVB3 variant H3N-375TS by inserting target sites (TS) of miR-375, which is specifically expressed in the pancreas, into the 3'UTR of the genome of the pancreo- and cardiotropic CVB3 variant H3. In vitro evaluation showed that H3N-375TS was suppressed in pancreatic miR-375-expressing EndoC-βH1 cells >5 log10, whereas its replication was not suppressed in isolated primary embryonic mouse cardiomyocytes. In vivo, intraperitoneal (i.p.) administration of H3N-375TS to NMRI mice did not result in pancreatic or cardiac infection. In contrast, intravenous (i.v.) administration of H3N-375TS to NMRI and Balb/C mice resulted in myocardial infection and acute and chronic myocarditis, whereas the virus was not detected in the pancreas and the pancreatic tissue was not damaged. Acute myocarditis was characterized by myocardial injury, inflammation with mononuclear cells, induction of proinflammatory cytokines, and detection of replicating H3N-375TS in the heart. Mice with chronic myocarditis showed myocardial fibrosis and persistence of H3N-375TS genomic RNA but no replicating virus in the heart. Moreover, H3N-375TS infected mice showed distinctly less suffering compared with mice that developed pancreatitis and myocarditis after i.p. or i.v application of control virus. CONCLUSION: In this study, we demonstrate that by use of the miR-375-sensitive CVB3 variant H3N-375TS, CVB3myocarditis can be established without the animals developing severe systemic infection and pancreatitis. As the H3N-375TS myocarditis model depends on pancreas-attenuated H3N-375TS, it can easily be used in different mouse strains and for various applications. Published on behalf of the European Society of Cardiology. All rights reserved.
Authors: Dirk M Nettelbeck; Mathias F Leber; Jennifer Altomonte; Assia Angelova; Julia Beil; Susanne Berchtold; Maike Delic; Jürgen Eberle; Anja Ehrhardt; Christine E Engeland; Henry Fechner; Karsten Geletneky; Katrin Goepfert; Per Sonne Holm; Stefan Kochanek; Florian Kreppel; Lea Krutzke; Florian Kühnel; Karl Sebastian Lang; Antonio Marchini; Markus Moehler; Michael D Mühlebach; Ulrike Naumann; Roman Nawroth; Jürg Nüesch; Jean Rommelaere; Ulrich M Lauer; Guy Ungerechts Journal: Viruses Date: 2021-07-21 Impact factor: 5.048
Authors: Ahmet Hazini; Babette Dieringer; Karin Klingel; Markian Pryshliak; Anja Geisler; Dennis Kobelt; Ole Daberkow; Jens Kurreck; Sophie van Linthout; Henry Fechner Journal: Viruses Date: 2021-09-24 Impact factor: 5.048