RESEARCH SUPPORT, NON-U.S. GOV'T
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A screening assay for the identification of host cell requirements and antiviral targets for hepatitis D virus infection.

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.

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