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Real-time quantitative PCR analysis of human HepaRG cells infected with hepatitis E virus (HEV) [D100 plaque 2]

ABSTRACT: We aimed to identify interferon (IFN)-regulated genes that are differentially expressed during chronic HEV infection in human hepatocytes, the main site of HEV replication, using HepaRG cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE243860 | GEO | 2023/11/08

REPOSITORIES: GEO

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Real-time quantitative PCR analysis of human HepaRG cells infected with hepatitis E virus (HEV) [D100 plaque 1]

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Real-time quantitative PCR analysis of human HepaRG cells infected with hepatitis E virus (HEV) [pre-array]

Project description:We aimed to identify interferon (IFN)-regulated genes that are differentially expressed during chronic HEV infection in human hepatocytes, the main site of HEV replication, using HepaRG cells. We have performed a preliminary screen using whole-cell RNA extracts prepared from HepaRG mock-infected or infected cells to determine whether HEV was able to trigger an IFN response. Different multiplicities of infection (MOI) (10 and 100 genome equivalent (GE)/cell) and time points (D+7, D+14, D+26, D+40, D+72 and D+100) were analyzed using the PCR array. We have also treated HepaRG cells after overnight treatment with IFN-β to confirm the ability of HepaRG to respond to IFN-I treatment.

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Real-time quantitative PCR analysis of liver tissue from pigs infected experimentally with hepatitis E virus (HEV) [pig liver]

Project description:We aimed to identify interferon (IFN)-regulated genes that are differentially expressed during HEV infection in swine liver tissues.

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Project description:Hepatitis E virus (HEV) is a globally prevalent pathogen that causes 20 million infections and 60,000 fatalities annually, endangering particularly pregnant women and immunosuppressed individuals. Liver cirrhosis, which results from advanced fibrosis, is the primary symptom and leading mortality cause in chronic hepatitis E patients. However, the causation and process of liver fibrosis triggered by chronic HEV infection remain poorly understood. Here, we unexpectedly discovered that the viral multiple-domain replicase (ORF1) undergoes unique ubiquitin-proteasomal processing in HEV replicon hepatocytes, HEV-infected gerbil livers, and HEV-infected patient livers, which follows a CHIP-mediated K48 ubiquitination and produces the HEV-Derived Smad Activator (HDSA). Lacking putative helicase and RNA polymerase domains, this enriched viral polypeptide in hepatocytes and gerbil livers is non-HSP90-bound, stable, and exhibits exclusively nuclear localization. Surprisingly, HDSA markedly potentiates the fibrogenic TGF-β/Smad pathway in livers by facilitating promoter binding and coactivator recruitment of SMAD3, leading to profound liver fibrotic symptoms and damage. Thus, we have identified the first viral protein derived from the unique proteasomal processing of the host, defined its notable role in liver fibrosis, and highlighted the nature of complex host-HEV interactions that drives HEV pathogenesis.

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Hepatocytes-HepaRG reprogramming.

Project description:To identify the molecular mechanisms and environmental inducers contributing to reprogramming of hepatocytes into progenitors in HCC context, we used the HepaRG cell line as model.

2013-12-05 | GSE52989 | GEO

Real-time quantitative PCR analysis of liver tissue from pigs infected experimentally with hepatitis E virus (HEV)

Project description:This SuperSeries is composed of the SubSeries listed below.

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Proteome and secretome of HepaRG cells

Project description:Primary human hepatocytes, the gold standard for in vitro studies of liver-related functions, suffer from uncertain availability and high variability in cell activity. Over years, a number of alternative hepatic cell lines have lost major liver-like functions, but not HepaRG cells. Therefore, their increasing use worldwide today arouses the need for establishing a reference functional status of differentiated HepaRG cells known as HPR116, which originate from the initial cell bank. Deep proteome and secretome analyses enabled us to show that they nicely express, at levels generally close to those in primary hepatocytes, master regulators of the hepatic phenotype, structural elements that ensure liver-like polarisation and factors supporting their transdifferentiation properties. Their highly differentiated status, mitochondrial functionality, hepatokine secretion ability and response to insulin was verified. Overall, the HepaRG cell system appears as robust surrogate cell system to primary hepatocytes, versatile enough to study not only xenobiotic detoxification but also the control of hepatic energy metabolism, secretory function and disease-related signalling pathways.

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HepaRG cells as a model of the primary human hepatocyte transcriptome

Project description:The aim of this experiment is to determine the similarities and differences between gene expression profiles in HepaRG cells versus primary human hepatocytes, human liver, and the commonly used HepG2 cell. We compared the gene expression profiles from replicate triplicate biological samples of human liver, primary human hepatocytes, HepG2 cells, and HepaRG at differing levels of maturity (differentiated and undifferentiated).

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