Skip navigation
Please use this identifier to cite or link to this item:
Title: Designing a system to investigate the impact of spatial transcriptional differences in the liver on hepatitis delta virus infection
Authors: Guo, Raymond
Advisors: Ploss, Alexander
Department: Molecular Biology
Class Year: 2019
Abstract: Hepatitis B virus (HBV) and the satellite hepatitis delta virus (HDV) cause liver infections which affect millions worldwide. HBV/HDV co-infection of human liver hepatocytes, which are important to metabolism, causes accelerated liver damage and elevated risk of hepatocellular carcinoma. Environmental cues affect hepatocytes’ transcriptional profile, and transcriptional differences divide the liver into zones. However, it is unclear if all hepatocytes are equally susceptible to HDV infection, and we hypothesize that liver zonation patterns distinctively affect permissiveness to viral infection. The Ploss Lab has generated a novel genetically humanized mouse model supporting HDV infection, and in order to utilize this mouse model to investigate the effects of liver zonation on HDV infection, we have designed a novel reporter system fusing the only gene product of HDV, the hepatitis delta antigen (HDAg), to Cre recombinase to enable permanent genetic marking of hepatocytes that take up this “HDVLP-Cre.” HDV-Cre DNA in vitro transduction experiments and hydrodynamic delivery of DNA plasmid to humanized mice confirm recombinase activity, and in vitro HDVLP-Cre infections of reporter cell lines demonstrate overall reporter activation. However, packaging of HDVLP-Cre proved to be difficult yielding relatively low reporter activity of the HDVLP-Cre construct which hampered more significant progress. Efforts to improve overall reporter activity included accelerating the experimental time frame as well as investigating alternative reporter cell lines, which yielded varying amounts of increased reporter activation. Moving forward, the production of HDVLP-Cre and overall HDV-Cre reporter construct will be further optimized for use in humanized mouse infections. Following in vivo infections of humanized mice, a potential unique zone of hepatocytes susceptible to initial HDV infection may be identified by analyzing hepatocytes which are permissive to HDVLP-Cre infection, allowing us to further investigate host factors which promote the establishment of HDV infection. The implications of this project can thus lead to future studies of targeted therapeutic treatment of HBV and HDV.
Type of Material: Princeton University Senior Theses
Language: en
Appears in Collections:Molecular Biology, 1954-2024

Files in This Item:
File Description SizeFormat 
GUO-RAYMOND-THESIS.pdf5.93 MBAdobe PDF    Request a copy

Items in Dataspace are protected by copyright, with all rights reserved, unless otherwise indicated.