Pan-Genotype Hepatitis E Virus Replication in Stem Cell-Derived Hepatocellular Systems.

Détails

ID Serval
serval:BIB_D1EA633DF71C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Pan-Genotype Hepatitis E Virus Replication in Stem Cell-Derived Hepatocellular Systems.
Périodique
Gastroenterology
Auteur(s)
Wu X., Dao Thi V.L., Liu P., Takacs C.N., Xiang K., Andrus L., Gouttenoire J., Moradpour D., Rice C.M.
ISSN
1528-0012 (Electronic)
ISSN-L
0016-5085
Statut éditorial
Publié
Date de publication
02/2018
Peer-reviewed
Oui
Volume
154
Numéro
3
Pages
663-674.e7
Langue
anglais
Notes
Publication types: Comparative Study ; Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
The 4 genotypes of hepatitis E virus (HEV) that infect humans (genotypes 1-4) vary in geographical distribution, transmission, and pathogenesis. Little is known about the properties of HEV or its hosts that contribute to these variations. Primary isolates grow poorly in cell culture; most studies have relied on variants adapted to cancer cell lines, which likely alter virus biology. We investigated the infection and replication of primary isolates of HEV in hepatocyte-like cells (HLCs) derived from human embryonic and induced pluripotent stem cells.
Using a cell culture-adapted genotype 3 strain and primary isolates of genotypes 1 to 4, we compared viral replication kinetics, sensitivity to drugs, and ability of HEV to activate the innate immune response. We studied HLCs using quantitative reverse-transcriptase polymerase chain reaction and immunofluorescence assay and enzyme-linked immunosorbent assays. We used an embryonic stem cell line that can be induced to express the CRISPR-Cas9 machinery to disrupt the peptidylprolyl isomerase A gene, encoding cyclophilin A (CYPA), a protein reported to inhibit replication of cell culture-adapted HEV. We further modified this line to rescue expression of CYPA before terminal differentiation to HLCs and performed HEV infection studies.
HLCs were permissive for infection by nonadapted, primary isolates of HEV genotypes 1 to 4. HEV infection of HLCs induced a replication-dependent type III interferon response. Replication of primary HEV isolates, unlike the cell culture-adapted strain, was not affected by disruption of the peptidylprolyl isomerase A gene or exposure to the CYPA inhibitor cyclosporine A.
Cell culture adaptations alter the replicative capacities of HEV. HLCs offer an improved, physiologically relevant, and genetically tractable system for studying the replication of primary HEV isolates. HLCs could provide a model to aid development of HEV drugs and a system to guide personalized regimens, especially for patients with chronic hepatitis E who have developed resistance to ribavirin.
Mots-clé
Antiviral Agents/pharmacology, Cell Differentiation, Cyclophilin A/genetics, Cyclophilin A/metabolism, Drug Resistance, Viral, Genotype, Hep G2 Cells, Hepatitis E virus/drug effects, Hepatitis E virus/genetics, Hepatitis E virus/growth & development, Hepatitis E virus/immunology, Hepatocytes/immunology, Hepatocytes/metabolism, Hepatocytes/virology, Host-Pathogen Interactions, Human Embryonic Stem Cells/immunology, Human Embryonic Stem Cells/metabolism, Human Embryonic Stem Cells/virology, Humans, Immunity, Innate, Induced Pluripotent Stem Cells/immunology, Induced Pluripotent Stem Cells/metabolism, Induced Pluripotent Stem Cells/virology, Kinetics, Phenotype, RNA, Viral/genetics, Sofosbuvir/pharmacology, Time Factors, Transfection, Virus Replication/drug effects, Antiviral, HLCs, Personalized Medicine, Primary Isolates
Pubmed
Web of science
Création de la notice
13/01/2018 11:41
Dernière modification de la notice
20/08/2019 16:52
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