Functional and Biochemical Characterization of Hepatitis C Virus (HCV) Particles Produced in a Humanized Liver Mouse Model

Calattini, Sara; Fusil, Floriane; Mancip, Jimmy; Thi, Viet Loan Dao; Granier, Christelle; Gadot, Nicolas; Scoazec, Jean‐Yves; Zeisel, Mirjam B.; Baumert, Thomas F.; Lavillette, Dimitri; Dreux, Marlène; Cosset, François‐Loïc

doi:10.1074/jbc.m115.662999

Cited by 29 publications

(42 citation statements)

References 67 publications

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“…A hallmark of HCV infectious particles is their tight connection with very-low-density lipoproteins (VLDL) and low-density lipoproteins (LDL), giving rise to a hybrid form of lipoviral particles (LVPs) with heterogeneous buoyant density (5)(6)(7)(8)(9)(10)(11)(12). Nonexchangeable apolipoprotein B (apoB) and several exchangeable apolipoproteins (apoE, apoA-I, and apoC-I) have been found on the LVP surface (13)(14)(15).…”

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confidence: 99%

Neglected but Important Role of Apolipoprotein E Exchange in Hepatitis C Virus Infection

Yang

Wang

Chi

et al. 2016

J Virol

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Hepatitis C virus (HCV) is a major cause of chronic liver disease, infecting approximately 170 million people worldwide. HCV assembly is tightly associated with the lipoprotein pathway. Exchangeable apolipoprotein E (apoE) is incorporated on infectious HCV virions and is important for infectious HCV virion morphogenesis and entry. Moreover, the virion apoE level is positively correlated with its ability to escape E2 antibody neutralization. However, the role of apoE exchange in the HCV life cycle is unclear. In this study, the relationship between apoE expression and cell permissiveness to HCV infection was assessed by infecting apoE knockdown and derived apoE rescue cell lines with HCV. Exchange of apoE between lipoproteins and HCV lipoviral particles (LVPs) was evaluated by immunoprecipitation, infectivity testing, and viral genome quantification. Cell and heparin column binding assays were applied to determine the attachment efficiency of LVPs with different levels of incorporated apoE. The results showed that cell permissiveness for HCV infection was determined by exogenous apoE-associated lipoproteins. Furthermore, apoE exchange did occur between HCV LVPs and lipoproteins, which was important to maintain a high apoE level on LVPs. Lipid-free apoE was capable of enhancing HCV infectivity for apoE knockdown cells but not apoE rescue cells. A higher apoE level on LVPs conferred more efficient LVP attachment to both the cell surface and heparin beads. This study revealed that exogenous apoE-incorporating lipoproteins from uninfected hepatocytes safeguarded the apoE level of LVPs for more efficient attachment during HCV infection. IMPORTANCEIn this study, a neglected but important role of apoE exchange in HCV LVP infectivity after virus assembly and release was identified. The data indicated that apoE expression level in uninfected cells is important for high permissiveness to HCV infection. Secreted apoE-associated lipoprotein specifically enhances infection of HCV LVPs. apoE exchange between HCV LVP and lipoproteins is important to maintain an adequate apoE level on LVPs for their efficient attachment to cell surface. These data defined for the first time an extracellular role of exchangeable apoE in HCV infection and suggested that exchangeable apolipoproteins reach a natural equilibrium between HCV LVPs and lipoprotein particles, which provides a new perspective to the understanding of the heterogeneity of HCV LVPs in composition. M ore than 170 million people worldwide are infected with hepatitis C virus (HCV). Up to 80% of infected individuals are unable to clear the virus, and persistent infections lead to a high risk of developing liver cirrhosis and hepatocellular carcinoma (1). Direct-acting antivirals (DAA) significantly improved treatment efficiency, but an effective vaccine for the control of new HCV infection is still needed due to the limited access to anti-HCV treatment. Moreover, the emergence of DAA-resistant viruses calls for alternative strategies to control viral breakthrough (2-4).A...

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confidence: 99%

Neglected but Important Role of Apolipoprotein E Exchange in Hepatitis C Virus Infection

Yang

Wang

Chi

et al. 2016

J Virol

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“…Differences between in vitro– and in vivo–derived HCV particles have been reported. 22 , 23 , 30 , 31 , 32 In vivo–derived particles tend to be of lighter density and more infectious than in vitro–derived particles 2 , 22 and this may be because there is no human liver cell culture model that properly recapitulates VLDL biogenesis. 8 To test if the human liver chimeric FNRG mice could be used to address some of these differences we first set out to characterize the lipoprotein profile in this model.…”

Section: Resultsmentioning

confidence: 99%

“… 21 It remains unclear if HCV assembly and particle composition truly mimic HCV as a lipoviroparticle in human circulation. Articles presenting experiments in which HCVcc was passaged from cell culture to animal models (chimpanzees or liver chimeric Alb-uPA mice) or liver chimeric FRG mice 22 , 30 only reported representative buoyant density profiles that originated from a single time point or from pooled samples harvested at different time points. In this study, we showed that HCV buoyant density and infectivity change over time and are influenced by dietary changes.…”

Section: Discussionmentioning

confidence: 99%

Analysis of Hepatitis C Virus Particle Heterogeneity in Immunodeficient Human Liver Chimeric fah-/- Mice

Andréo

Jong

Scull

et al. 2017

Cellular and Molecular Gastroenterology and Hepatology

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Background & AimsHepatitis C virus (HCV) is a leading cause of chronic liver diseases and the most common indication for liver transplantation in the United States. HCV particles in the blood of infected patients are characterized by heterogeneous buoyant densities, likely owing to HCV association with lipoproteins. However, clinical isolates are not infectious in vitro and the relative infectivity of the particles with respect to their buoyant density therefore cannot be determined, pointing to the need for better in vivo model systems.MethodsTo analyze the evolution of the buoyant density of in vivo–derived infectious HCV particles over time, we infected immunodeficient human liver chimeric fumaryl acetoacetate hydrolase-/- mice with J6/JFH1 and performed ultracentrifugation of infectious mouse sera on isopicnic iodixanol gradients. We also evaluated the impact of a high sucrose diet, which has been shown to increase very-low-density lipoprotein secretion by the liver in rodents, on lipoprotein and HCV particle characteristics.ResultsSimilar to the severe combined immunodeficiency disease/Albumin-urokinase plasminogen activator human liver chimeric mouse model, density fractionation of infectious mouse serum showed higher infectivity in the low-density fractions early after infection. However, over the course of the infection, viral particle heterogeneity increased and the overall in vitro infectivity diminished without loss of the human liver graft over time. In mice provided with a sucrose-rich diet we observed a minor shift in HCV infectivity toward lower density that correlated with a redistribution of triglycerides and cholesterol among lipoproteins.ConclusionsOur work indicates that the heterogeneity in buoyant density of infectious HCV particles evolves over the course of infection and can be influenced by diet.

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“…In vivo models of HCV infection are also questionable, since small rodents cannot be infected with human HCV, and they only transgenically express HCV proteins, in the absence of replicating virus. The best option in this case is to use immunodeficient mice with humanized livers that are able to replicate the human virus, but to our best knowledge, while infection of the liver with HCV is widely reported (5,26,57), HCV-ethanol studies on this expensive and demanding model are very few (37).…”

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Role of apoptotic hepatocytes in HCV dissemination: regulation by acetaldehyde

Ganesan

Natarajan

Zhang

et al. 2016

American Journal of Physiology-Gastrointestinal and Liver Physi

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Alcohol consumption exacerbates hepatitis C virus (HCV) pathogenesis and promotes disease progression, although the mechanisms are not quite clear. We have previously observed that acetaldehyde (Ach) continuously produced by the acetaldehyde-generating system (AGS), temporarily enhanced HCV RNA levels, followed by a decrease to normal or lower levels, which corresponded to apoptosis induction. Here, we studied whether Ach-induced apoptosis caused depletion of HCV-infected cells and what role apoptotic bodies (AB) play in HCV-alcohol crosstalk. In liver cells exposed to AGS, we observed the induction of miR-122 and miR-34a. As miR-34a has been associated with apoptotic signaling and miR-122 with HCV replication, these findings may suggest that cells with intensive viral replication undergo apoptosis. Furthermore, when AGS-induced apoptosis was blocked by a pan-caspase inhibitor, the expression of HCV RNA was not changed. AB from HCV-infected cells contained HCV core protein and the assembled HCV particle that infect intact hepatocytes, thereby promoting the spread of infection. In addition, AB are captured by macrophages to switch their cytokine profile to the proinflammatory one. Macrophages exposed to HCV ϩ AB expressed more IL-1␤, IL-18, IL-6, and IL-10 mRNAs compared with those exposed to HCV Ϫ AB. The generation of AB from AGS-treated HCV-infected cells even enhanced the induction of aforementioned cytokines. We conclude that HCV and alcohol metabolites trigger the formation of AB containing HCV particles. The consequent spread of HCV to neighboring hepatocytes via infected AB, as well as the induction of liver inflammation by AB-mediated macrophage activation potentially exacerbate the HCV infection course by alcohol and worsen disease progression. HCV RNA; acetaldehyde; apoptosis; hepatocytes; macrophages ALCOHOL EXPOSURE EXACERBATES hepatitis C virus (HCV) infection severity, increases liver inflammation, and potentiates liver injury progression. However, the pathogenesis of HCV-alcohol interactions is not clear yet. The major difficulty in HCValcohol studies is related to the lack of adequate models that can recapitulate both viral replication and ethanol metabolism-two important features that potentiate liver injury progression in alcohol-abusing HCV patients. Human liver cell lines permissive for HCV (Huh7.5 or Huh 7.5.1) that are currently used for HCV-alcohol-related in vitro studies do not metabolize ethanol, while ethanol-metabolizing rodent liver cells cannot support HCV replication. Studies conducted with isolated human hepatocytes also have failed, since these cells require at least 4 -5 days to become infected with HCV; however, by this time, hepatocytes lose the expression and functional activity of CYP2E1 and alcohol dehydrogenase (ADH), the main ethanol-metabolizing enzymes (47). In vivo models of HCV infection are also questionable, since small rodents cannot be infected with human HCV, and they only transgenically express HCV proteins, in the absence of replicating virus. The best option i...

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Functional and Biochemical Characterization of Hepatitis C Virus (HCV) Particles Produced in a Humanized Liver Mouse Model

Cited by 29 publications

References 67 publications

Neglected but Important Role of Apolipoprotein E Exchange in Hepatitis C Virus Infection

Neglected but Important Role of Apolipoprotein E Exchange in Hepatitis C Virus Infection

Analysis of Hepatitis C Virus Particle Heterogeneity in Immunodeficient Human Liver Chimeric fah-/- Mice

Role of apoptotic hepatocytes in HCV dissemination: regulation by acetaldehyde

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