In vitro infection of primary human hepatocytes by HCV-positive sera: insights on a highly relevant model

Gondeau, Claire; Briolotti, Philippe; Razafy, Francia; Duret, Cédric; Rubbo, Pierre-Alain; Helle, François; Rème, Thierry; Ripault, Marie–Pierre; Ducos, J; Fabre, Jean‐Michel; Ramos, Jeanne; Pécheur, Eve‐Isabelle; Larrey, Dominique; Maurel, Patrick; Daujat-Chavanieu, Martine

doi:10.1136/gutjnl-2013-304623

Cited by 19 publications

(20 citation statements)

References 44 publications

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“…Moreover, APC expression in cancer cells causes nuclear and cytoplasmic translocation of b-catenin to the cell membrane (Aoki and Taketo, 2007). The increase of APC mRNA expression in parallel with PHHs adhesion and polarization (Gondeau et al, 2014) could contribute to WNT/b-catenin pathway down-regulation in PHHs (Hanson and Miller, 2005).…”

Section: Discussionmentioning

confidence: 99%

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

et al. 2014

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The wingless-type MMTV integration site family (WNT)/b-catenin/ adenomatous polyposis coli (CTNNB1/APC) pathway has been identified as a regulator of drug-metabolizing enzymes in the rodent liver. Conversely, little is known about the role of this pathway in drug metabolism regulation in human liver. Primary human hepatocytes (PHHs), which are the most physiologically relevant culture system to study drug metabolism in vitro, were used to investigate this issue. This study assessed the link between cytochrome P450 expression and WNT/b-catenin pathway activity in PHHs by modulating its activity with recombinant mouse Wnt3a (the canonical activator), inhibitors of glycogen synthase kinase 3b, and small-interfering RNA to invalidate CTNNB1 or its repressor APC, used separately or in combination. We found that the WNT/ b-catenin pathway can be activated in PHHs, as assessed by universal b-catenin target gene expression, leucine-rich repeat containing G protein-coupled receptor 5. Moreover, WNT/ b-catenin pathway activation induces the expression of CYP2E1, CYP1A2, and aryl hydrocarbon receptor, but not of CYP3A4, hepatocyte nuclear factor-4a, or pregnane X receptor (PXR) in PHHs. Specifically, we show for the first time that CYP2E1 is transcriptionally regulated by the WNT/b-catenin pathway. Moreover, CYP2E1 induction was accompanied by an increase in its metabolic activity, as indicated by the increased production of 6-OH-chlorzoxazone and by glutathione depletion after incubation with high doses of acetaminophen. In conclusion, the WNT/b-catenin pathway is functional in PHHs, and its induction in PHHs represents a powerful tool to evaluate the hepatotoxicity of drugs that are metabolized by CYP2E1.

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Section: Discussionmentioning

confidence: 99%

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

et al. 2014

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“…A multiplex profiling of cytokines and growth factors in serum samples from HCV-infected patients has shown that at least 24 cytokines and growth factors affect the change in hepatocyte biology and the efficiency of the infection process [4]. At the same time, only limited data are available for the roles particular regulators of immune reactions play in hepatitis C at the cell level (in infected liver cells).…”

Section: Introductionmentioning

confidence: 99%

Effect of Hepatitis C virus proteins on the production of proinflammatory and profibrotic cytokines in Huh7.5 human hepatoma cells

et al. 2016

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Hepatitis C virus (HCV) is a widespread dangerous human pathogen. Up to 80% of HCV-infected individuals develop chronic infection, which is often accompanied by liver inflammation and fibrosis and, at terminal stages, liver cirrhosis and cancer. Treatment of patients with end-stage liver disease is often ineffective, and even patients with suppressed HCV replication have higher risk of death as compared with noninfected subjects. Therefore, investigating the mechanisms that underlie HCV pathogenesis and developing treatments for virus-associated liver dysfunction remain an important goal. The effect of individual HCV proteins on the production of proinflammatory and profibrotic cytokines in hepatocellular carcinoma Huh7.5 cells was analyzed in a systematic manner. Cells were transfected with plasmids encoding HCV proteins. Cytokine production and secretion was accessed by immunocytochemistry and ELISA of the culture medium, and transcription of the cytokine genes was assessed using reverse transcription and PCR. HCV proteins proved to differ in effect on cytokine production. Downregulation of interleukin 6 (IL-6) production was observed in cells expressing the HCV core, NS3, and NS5A proteins. Production of transforming growth factor β1 (TGF-β1) was lower in cells expressing the core proteins, NS3, or E1/E2 glycoproteins. A pronounced increase in production and secretion of tumor necrosis factor α (TNF-α) was observed in response to expression of the HCV E1/E2 glycoproteins. A higher biosynthesis, but a lower level in the cell culture medium, was detected for interleukin 1β (IL-1β) in cells harboring NS4 and IL-6 in cells expressing NS5В. The finding was possibly explained by protein-specific retention and consequent accumulation of the respective cytokines in the cell.

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“…PHHs isolated from patient biopsies have been used to study HCV infection in vitro [53,146–152]. However, these cells suffer from limited accessibility, variability between batches, a finite culture time and low levels of infection with the standard HCVcc strains.…”

Section: Keeping the Pieces In Place: Role Of The Tight Junction In Ementioning

confidence: 99%

The Missing Pieces of the HCV Entry Puzzle

Ogden

Tang

2015

Future Virol.

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The past decade has witnessed steady and rapid progress in HCV research, which has led to the recent breakthrough in therapies against this significant human pathogen. Yet a deeper understanding of the life cycle of the virus is required to develop more affordable treatments and to advance vaccine design. HCV entry presents both a challenge for scientific research and an opportunity for alternative intervention approaches, owning to its highly complex nature and the myriad of players involved. More than half a dozen cellular proteins are implicated in HCV entry; and a more definitive picture regarding the structures of the glycoproteins is emerging. A role of apolipoproteins in HCV entry has also been established. Still, major questions remain, and the answers to these, which we summarize in this review, will hopefully close the gaps in our understanding and complete the puzzle that is HCV entry.

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In vitro infection of primary human hepatocytes by HCV-positive sera: insights on a highly relevant model

Cited by 19 publications

References 44 publications

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

Effect of Hepatitis C virus proteins on the production of proinflammatory and profibrotic cytokines in Huh7.5 human hepatoma cells

The Missing Pieces of the HCV Entry Puzzle

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