Blanchet, Matthieu; Angelo, Léna; Tetreault, Yasmine; Khabir, Marwa; Sureau, Camille; Vaillant, Andrew et Labonté, Patrick 
ORCID: https://orcid.org/0000-0001-7262-3125
(2023).
An in Vitro Culture Model for The Induction of HDV Replication in a HBV Expressing Cell Line
In: Meeting of the American-Association-for-the-Study-of-Liver-Diseases (AASLD), November 10-14, 2023, Boston, MA.
Résumé
Background: Individual’s chronically infected with
both Hepatitis B virus (HBV) and Hepatitis D virus
(HDV) present an increased risk to develop cirrhosis
and hepatocellular carcinoma in comparison to HBV
mono-infected individual’s. The number of HDV
infected individual’s worldwide is estimated to be
between 20 and 40 million, a strong incentive to
further engage in the search for new antivirals using
innovative and practical in vitro models. Here we
present the HepG2BD cell line as a novel in vitro
culture system for inducible HDV replication in HBV
replicating cells. Methods: Engineering: The
HepG2BD cell line derives from HepG2.2.15 cells in
which a HDV cDNA sequence under the control of a
Tet-Off promoter is inserted into the adeno-associated
virus integration site 1 (AAVS1) safe harbor site. The
insertion at this precise location was conducted using
the CRISPR-Cas9 technology for reliable initiation/
repression of HDV RNA replication, and production of infectious HDV virions in a cell line that constitutively
expresses HBV. Characterization: HDV RNA in cells
and supernatants was quantified by RT-qPCR and
Northern blot, HBV RNA in cells was monitored by RTqPCR. Secreted HBV particle concentrations were
evaluated by qPCR after anti-PreS1 immunoprecipitation. Secreted HBsAg was quantified by
ELISA. Cellular presence and localization of HDAg in
various cell culture conditions was monitored by
confocal immunofluorescence. Infectivity of secreted
HDV virions was measured by inoculation of NTCPHuh7 cells followed by RT-qPCR. Results: The
presented cell line is, to our knowledge, the first one
to allow full replication of both HBV and HDV. This cell
line shows proper induction of the HDV lifecycle. The
interplay between HBV and HDV was detected in
many instances. Differentiation into more classic
hepatocyte morphology was associated with an
increased concentration of both HDV and HBV in the
supernatant. Conclusion: This new and unique model
will be instrumental for the screening/characterization
of new antivirals targeting both viruses (such as REP
2139). The HepG2BD cell line is also expected to help
in our understanding of the dynamic interplay between
the HBV and HDV lifecycles, and in identifying cellular
factors involved.
| Type de document: | Document issu d'une conférence ou d'un atelier |
|---|---|
| Informations complémentaires: | Présentation orale 1200-C Hepatology 78 (Suppl.1): S367 http://dx.doi.org/ 10.1097/HEP.0000000000000580 |
| Mots-clés libres: | - |
| Centre: | Centre INRS-Institut Armand Frappier |
| Date de dépôt: | 06 janv. 2024 22:31 |
| Dernière modification: | 06 janv. 2024 22:31 |
| URI: | https://espace.inrs.ca/id/eprint/13965 |
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