Cellular models for the screening and development of anti-hepatitis C virus agents

Gondeau, Claire; Pichard-Garcia, Lydiane; Maurel, Patrick

doi:10.1016/j.pharmthera.2009.05.010

Cited by 22 publications

(16 citation statements)

References 337 publications

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“…In the past few years, robust cellular model systems that support HCV infection, replication, and viral particle secretion have been developed (27). Using these systems, studies have shown that HCV, like other positive-strand RNA viruses, hijacks intracellular membranes, probably of diverse origins, to generate unique membranous platforms where HCV genome replication and viral particle assembly occur (20).…”

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confidence: 99%

Role for ADP Ribosylation Factor 1 in the Regulation of Hepatitis C Virus Replication

Matto

Sklan

David

et al. 2011

J Virol

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We hypothesized that ADP-ribosylation factor 1 (Arf1) plays an important role in the biogenesis and maintenance of infectious hepatitis C virus (HCV). Huh7.5 cells, in which HCV replicates and produces infectious viral particles, were exposed to brefeldin A or golgicide A, pharmacological inhibitors of Arf1 activation. Treatment with these agents caused a reduction in viral RNA levels, the accumulation of infectious particles within the cells, and a reduction in the levels of these particles in the extracellular medium. Fluorescence analyses showed that the viral nonstructural (NS) proteins NS5A and NS3, but not the viral structural protein core, shifted their localization from speckle-like structures in untreated cells to the rims of lipid droplets (LDs) in treated cells. Using pulldown assays, we showed that ectopic overexpression of NS5A in Huh7 cells reduces the levels of GTP-Arf1. Downregulation of Arf1 expression by small interfering RNA (siRNA) decreased both the levels of HCV RNA and the production of infectious viral particles and altered the localization of NS5A to the peripheries of LDs. Together, our data provide novel insights into the role of Arf1 in the regulation of viral RNA replication and the production of infectious HCV.

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confidence: 99%

Role for ADP Ribosylation Factor 1 in the Regulation of Hepatitis C Virus Replication

Matto

Sklan

David

et al. 2011

J Virol

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“…Some candidate anti-viral molecules which first require metabolic activation (e.g., phosphorylation) to function properly in vivo may test negative in the cultured cell lines due to the lack of pre-activation in vitro. On the other hand, some candidate molecules that screen positive in cultured cell lines may not exert antiviral activities in vivo due to being metabolized before they are able to function in target tissues [20]. Because of the lack of a suitable small animal model, only two NS3/4A-specific inhibitors have advanced into clinical applications at present [8,9].…”

Section: Discussionmentioning

confidence: 99%

Establishment of a novel triple-transgenic mouse: conditionally and liver-specifically expressing hepatitis C virus NS3/4A protease

et al. 2014

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It is well known that NS3/4A protein plays crucial roles in the hepatitis C virus (HCV) replication. NS3/4A protein also results to virus-mediated immune evasion and persistence of infection through the interaction with host proteins. However, the lack of a suitable animal model hampers studies of HCV NS3/4A protein interaction with host proteins, which impacts immunopathology due to infection. Here, transgenic vector containing transcriptional regulation and Fluc reporter gene was constructed to conditionally express NS3/4A protein under the dual control of Tet-On regulatory system and Cre/LoxP gene-knockout system. NS3/4A transgenic founder mice were continuously crossed with Lap transgenic mice expressing reverse tetracycline-controlled transcriptional activator (rtTA), the NS3/4A/Lap double transgenic mouse lines with liver-specifically and conditionally expressing reporter (luciferase Fluc) under control of Tet-On system were established. The NS3/4A/Lap double transgenic mouse are mated with Lap/LC-1 double transgenic mouse with liver-specifically and conditionally expressing Cre recombinase under control of Tet-On system, NS3/4A/Lap/LC-1 triple transgenic mouse were generated. In vivo bioluminescent imaging, western blotting and immunohistochemical staining (IHS) was used to confirm that NS3/4A protein was strictly expressed in the liver of Doxycycline-induced triple transgenic mice. The results show that we established a triple-transgenic mouse model conditionally expressing the HCV NS3/4A protein under strict control of the Tet-On regulatory system and Cre/loxP system. This novel transgenic mouse model expressing NS3/4A in a temporally and spatially-specific manner will be useful for studying interactions between HCV NS3/4A protein and the host, also for evaluating NS3/4A protease inhibitors.

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“…Post-binding steps are followed by a lateral migration, and the interaction of the virus with claudin-1 and occludin that leads to the tight junctions integrity disruption and culminate in clathrin-dependent endocytosis of virus particles, their endosome membrane fusion and viral genome release. facilitated the discovery and characterization of new receptors involved in virus-cell attachment and entry [39][40][41][42] other than the CD81 and LDL-receptor [43][44][45], enabling the development of new viral targets and therapies [46].…”

Section: Virus Attachment and Penetrationmentioning

confidence: 99%

HCV Infection by Cell-to-Cell Transmission: Choice or Necessity?

Carloni

Crema²,

Valli

et al. 2012

CMM

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In vitro models of HCV infection have allowed for the clarifying of molecules and mechanisms involved in the main steps of virus cell-entry. HCV entry and neutralization appear to be closely related. Neutralizing antibodies inhibit the E2-CD81 binding, therefore CD81 is considered to be a major target of immune response. The tight-junction proteins are also implicated in E2-binding to CD81 and successive steps of virus entry, in cooperation with several co-receptors, whose involvement has still to be elucidated. Increasing evidence has emphasized the importance of cell-to-cell HCV-transmission in chronic infection. This route for infection could favour virus-escape from host-neutralization though its CD81-dependency is still debated. The main reasons which have delayed our understanding of HCV-infection are here critically reviewed, as are the challenges faced by investigators in the field. A deeper insight into the different pathways involved could help to elucidate some crucial features of HCV infection mechanisms and disclose important implications in its pathogenesis, which could help in suggesting new targets for successful immune-prophylactic/therapeutic strategies.

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Cellular models for the screening and development of anti-hepatitis C virus agents

Cited by 22 publications

References 337 publications

Role for ADP Ribosylation Factor 1 in the Regulation of Hepatitis C Virus Replication

Role for ADP Ribosylation Factor 1 in the Regulation of Hepatitis C Virus Replication

Establishment of a novel triple-transgenic mouse: conditionally and liver-specifically expressing hepatitis C virus NS3/4A protease

HCV Infection by Cell-to-Cell Transmission: Choice or Necessity?

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