GOLT1B Activation in Hepatitis C Virus-Infected Hepatocytes Links ER Trafficking and Viral Replication.

Jacqueline Butterworth,Edouard Bertrand,Alessia Virzì,Guillaume Desandré,Eric Assenat,Urszula Hibner,Jean-Christophe Andrau,Marion Peter,Armando Andres Roca Suarez,Yannick Simonin,Marine Guivarch,Amal Zine El Aabidine,Damien Gregoire,Joachim Lupberger

doi:10.3390/pathogens11010046

Abstract

Chronic hepatitis C carries a high risk of development of hepatocellular carcinoma (HCC), triggered by both direct and indirect effects of the virus. We examined cell-autonomous alterations in gene expression profiles associated with hepatitis C viral presence. Highly sensitive single molecule fluorescent in situ hybridization applied to frozen tissue sections of a hepatitis C patient allowed the delineation of clusters of infected hepatocytes. Laser microdissection followed by RNAseq analysis of hepatitis C virus (HCV)-positive and -negative regions from the tumoral and non-tumoral tissues from the same patient revealed HCV-related deregulation of expression of genes in the tumor and in the non-tumoral tissue. However, there was little overlap between both gene sets. Our interest in alterations that increase the probability of tumorigenesis prompted the examination of genes whose expression was increased by the virus in the non-transformed cells and whose level remained high in the tumor. This strategy led to the identification of a novel HCV target gene: GOLT1B, which encodes a protein involved in ER-Golgi trafficking. We further show that GOLT1B expression is induced during the unfolded protein response, that its presence is essential for efficient viral replication, and that its expression is correlated with poor outcome in HCC.

Highlights

Chronic hepatitis C predisposes patients to life-threatening pathologies, including hepatocellular carcinoma (HCC) [1]
In order to further test the sensitivity of our detection procedure, we explored detection of the signal in frozen liver sections from a hepatitis C virus (HCV) transgenic FL-N/35 mouse model, expressing very low levels of viral RNA [24] and reminiscent of the clinically relevant situation found in livers of chronically infected hepatitis C patients
In addition to necro-inflammatory liver damage, characteristic of chronic viral hepatitis and responsible for creating a favorable environment for tumor development, direct effects of the virus on the host cell have been incriminated as HBV-induced pro-oncogenic events

Summary

Introduction

Chronic hepatitis C predisposes patients to life-threatening pathologies, including hepatocellular carcinoma (HCC) [1]. The persistence of the virus and the accompanying inflammation-driven liver disease are major contributors to the progression from steatosis to fibrosis, cirrhosis, and HCC. Cell-autonomous effects of the virus that disturb cellular homeostasis and generate a pro-oncogenic environment via deregulation of hepatic metabolism and signal transduction further increase the risk of tumorigenesis (for review see [2,3,4]). In the case of hepatitis C virus (HCV), the distinction between direct and indirect effects of the virus on the host cell is not clear-cut. An unbiased genome-wide proteogenomic approach highlighted major HCC hallmarks that are induced by HCV infection, including EGFR, STAT3, epithelial–mesenchymal transition (EMT), and perturbations in liver metabolism and DNA repair [10,11]. Despite recent progress in our understanding of the virus–host interactions, much remains to be learned about subtle virus-driven cell-autonomous alterations that may escape detection in a bulk analysis of the “omics” landscape

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Results

Conclusion