title: A screening assay for the identification of host cell requirements and antiviral targets for hepatitis D virus infection.
abstract: Hepatitis delta virus (HDV) is a minimalistic satellite virus of hepatitis B virus (HBV). HBV/HDV co-infection, i.e. "hepatitis D", is the most severe form of viral hepatitis. No effective therapy for HDV infection is available partly due to the fact that HDV is a highly host-dependent virus devoid of any potentially drugable enzyme encoded in its small genome. In this study we present a semi-automated method to evaluate HDV infection and replication under the influence of different drugs. We utilized a Huh-7/hNTCP cell culture based system in a 96-well plate format, an automated microscope and image acquisition as well as analysis with the CellProfiler software to quantify the impact of these drugs on HDV infection. For validation, three groups of potential anti-HDV agents were evaluated: To target ribozyme activity of HDV RNA, we screened ribozyme inhibitors but only observed marked toxicity. Testing innate antiviral mediators showed that interferons alpha-2a and beta-1a had a specific inhibitory effect on HDV infection. Finally, we screened a library of 160 human kinase inhibitors covering all parts of the human kinome. Overall, only inhibitors targeting the tyrosine kinase-like group had significant average anti-HDV activity. Looking at individual substances, kenpaullone, a GSK-3β and Cdk inhibitor, had the highest selective index of 3.44. Thus, we provide a potentially useful tool to screen for substances with anti-HDV activity and novel insights into interactions between HDV replication and the human kinome.
authors.affiliation: Institute for Molecular Biology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Buchmann
authors.affiliation: Institute of Virology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany.
authors.forename: Katinka
authors.initials: K
authors.lastname: Döhner
authors.affiliation: Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schirdewahn
authors.affiliation: German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany; Institute of Virology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Sodeik
authors.affiliation: Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Manns
authors.affiliation: Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany.
authors.forename: Heiner
authors.initials: H
authors.lastname: Wedemeyer
authors.affiliation: Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany; Institute of Virology, University Hospital Essen, University of Duisburg- Essen, D-45147 Essen, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Ciesek
authors.affiliation: Institute for Molecular Biology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl- Neuberg Str.1, D-30625 Hannover, Germany; German Center for Infection Research (DZIF), Braunschweig-Hannover site, Germany. Electronic address: vonhahn.thomas@mh-hannover.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: von Hahn
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000998:Antiviral Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001331:Automation; D001552:Benzazepines / Q000494:pharmacology; D060085:Coinfection / Q000188:drug therapy; D006508:Hepatitis Antibodies / Q000276:immunology*; D006509:Hepatitis B / Q000821:virology; D006515:Hepatitis B virus / Q000302:isolation & purification; D003699:Hepatitis D / Q000188:drug therapy* / Q000821:virology; D003698:Hepatitis Delta Virus / Q000187:drug effects*; D022781:Hepatocytes / Q000187:drug effects / Q000821:virology; D057166:High-Throughput Screening Assays / Q000379:methods*; D054884:Host-Pathogen Interactions / Q000187:drug effects*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016337:RNA, Catalytic / Q000037:antagonists & inhibitors; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D001552:Benzazepines; D006508:Hepatitis Antibodies; D007211:Indoles; D047428:Protein Kinase Inhibitors; D016337:RNA, Catalytic; C119620:kenpaullone; D011505:Protein-Tyrosine Kinases
keywords: Automated screen; Hepatitis delta virus (HDV); Human kinome; Kinase inhibitors