Principal Investigator: Ass Prof. Nicola Fletcher, University College Dublin, Ireland
The growth of hepatitis E virus (HEV) in culture has proven to be very challenging, and most studies have used cell culture-adapted viral strains and transformed cell lines. The lack of a robust cell culture model for the study of HEV infection is one of the greatest roadblocks to our understanding of factors that determine HEV transmission and to the development of novel anti-viral treatments for HEV. Microscopy analysis of 3D cell cultures and tissue organoids is challenging because it combines mutually exclusive requirements.
CoCID employs SXM to bridge the current gap between light and electron microscopy and will serve as an interface for correlative studies. A SXM with high spatial resolution in 3D method allows for volume rendering of cells in a few minutes. We will visualize the sites of HEV as well as viral replication and assembly in pig and human polarized and non-polarised cell lines and tissues.
Furthermore, we will investigate the ability of a panel of inhibitors and antiviral agents to reverse observed structural changes, and perform parallel infectivity assays to uncover novel antiviral targets.
Due to the lack of in vitro model systems that support robust HEV infection, there are few studies that have attempted to elucidate the steps of infection, i.e. viral entry, replication and assembly. Here we will use recently developed model systems for HEV that recapitulate the unique polarized nature of hepatocytes to study HEV replication in human and pig hepatocytes. This study will allow analysis of full lifecycle of HEV in human and pig liver models and to image the effect of antiviral therapeutics and inhibitors on infection.
SXM imaging and data analysis will bridge the gap between light and electron microscopy that have previously been applied, and will serve as an interface for correlative studies. We will visualize sites of HEV replication and assembly in pig and human polarized and non-polarised cell lines, and confirm our findings using SXM combined with other imaging techniques such as EM and confocal microscopy.