Dual Effects of Let-7b in the Early Stage of Hepatitis C Virus Infection

MicroRNA let-7b expression is induced by infection of hepatitis C virus (HCV) and is involved in the regulation of HCV replication by directly targeting the HCV genome. The current study demonstrated that let-7b directly targets negative regulators of type I interferon (IFN) signaling thereby limiting HCV replication in the early stage of HCV infection. Let-7b-regulated genes which are involved in host cellular responses to HCV infection were unveiled by microarray profiling and bioinformatic analyses followed by various molecular and cellular assays using Huh7 cells expressing wild type or the seed region-mutated let-7b. Let-7b targeted the cytokine signaling 1 (SOCS1) protein, a negative regulator of JAK/STAT signaling, which then enhanced STAT1-Y701 phosphorylation leading to increased expression of the downstream interferon stimulated genes (ISGs). Let-7b augmented RIG-I signaling, but not MDA5, to phosphorylate and nuclear translocate IRF3 leading to increased expression of IFN-β. Let-7b directly targeted the ATG12 and IKKα transcripts and reduced the interaction of the ATG5-ATG12 conjugate and RIG-I leading to increased expression of IFN, which may further stimulate JAK/STAT signaling. Let-7b induced by HCV infection elicits dual effects on IFN expression and signaling, along with targeting the coding sequences of NS5B and 5'-UTR of HCV genome, limited HCV RNA accumulation in the early stage of HCV infection. Controlling let-7b expression is thereby crucial in the intervention of HCV infection. Importance: HCV is a leading cause of liver disease, with an estimated 71 million people infected worldwide. During HCV infection, type I IFN signaling displays potent anti-viral and immuno-modulatory effects. Host factors, including microRNAs, play a role in up-regulating IFN signaling to limit HCV replication. Let-7b is a liver abundant miRNA that is induced by HCV infection and targets the HCV genome to suppress HCV RNA accumulation. In this study, we demonstrated that let-7b, as a positive regulator of type I IFN signaling, plays dual roles in against HCV replication by increasing the expression of IFN and ISRE-driven ISGs in the early stage of HCV infection. This study sheds new insight into understanding the role of let-7b in combatting HCV infection. Clarifying IFN signaling regulated by miRNA during the early phase of HCV infection may help researchers understand the initial defense mechanisms to other RNA viruses.

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Inhibition of STAT Pathway Impairs Anti-Hepatitis C Virus Effect of Interferon Alpha

Cellular Physiology and Biochemistry ◽

10.1159/000452526 ◽

2016 ◽

Vol 40 (1-2) ◽

pp. 77-90 ◽

Cited By ~ 7

Author(s):

Lan-Juan Zhao ◽

Sheng-Fei He ◽

Yuan Liu ◽

Ping Zhao ◽

Zhong-Qi Bian ◽

...

Keyword(s):

Gene Expression ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Interferon Alpha ◽

Hcv Infection ◽

Hcv Rna ◽

Jak Inhibitor ◽

Stat Signaling ◽

Antiviral Gene ◽

Stat Pathway

Background/Aims: Signal transducer and activator of transcription (STAT) pathway plays an important role in antiviral efficacy of interferon alpha (IFN-α). IFN-α is the main therapeutic against hepatitis C virus (HCV) infection. We explored effects of IFN-α on HCV replication and antiviral gene expression by targeting STAT. Methods: In response to IFN-α, STAT status, HCV replication, and antiviral gene expression were analyzed in human hepatoma Huh7.5.1 cells before and after cell culture-derived HCV infection. Results: IFN-α treatment induced expression and phosphorylation of STAT1 and STAT2 in Huh7.5.1 cells. Pretreatment of Huh7.5.1 cells with a mAb to IFN alpha receptor (IFNAR) 2 decreased IFN-α-dependent phosphorylation of STAT1 and STAT2, whereas pretreatment with an IFNAR1 mAb increased such phosphorylation, suggesting that IFNAR mediates IFN-α-triggered STAT signaling. During HCV infection, STAT1 and STAT2 phosphorylation could be rescued by IFN-α and IFN-α-induced phosphorylation of STAT1 and STAT2 was impaired. Inhibition of STAT pathway by Jak inhibitor I significantly enhanced HCV RNA replication and viral protein expression. Antiviral genes coding for IFN regulatory factor 9 and IFN-stimulated gene 15 were up-regulated by IFN-α during HCV infection but such up-regulation was abrogated by Jak inhibitor I. Conclusion: These results establish that activation of STAT pathway is essential for anti-HCV efficacy of IFN-α. Impairment of IFN-α-triggered STAT signaling by HCV may account for evading IFN-α response.

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Replication of Subgenomic Hepatitis C Virus Replicons in Mouse Fibroblasts Is Facilitated by Deletion of Interferon Regulatory Factor 3 and Expression of Liver-Specific MicroRNA 122

Journal of Virology ◽

10.1128/jvi.00559-10 ◽

2010 ◽

Vol 84 (18) ◽

pp. 9170-9180 ◽

Cited By ~ 54

Author(s):

Liang-Tzung Lin ◽

Ryan S. Noyce ◽

Tram N. Q. Pham ◽

Joyce A. Wilson ◽

Gary R. Sisson ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Mouse Models ◽

Type I Interferon ◽

Hcv Infection ◽

Host Factors ◽

Hcv Rna ◽

Type I ◽

Interferon Signaling ◽

Mouse Cells

ABSTRACT Hepatitis C virus (HCV) infection causes significant morbidity, and efficient mouse models would greatly facilitate virus studies and the development of effective vaccines and new therapeutic agents. Entry factors, innate immunity, and host factors needed for viral replication represent the initial barriers that restrict HCV infection of mouse cells. Experiments in this paper consider early postentry steps of viral infection and investigate the roles of interferon regulatory factors (IRF-3 and IRF-9) and microRNA (miR-122) in promoting HCV replication in mouse embryo fibroblasts (MEFs) that contain viral subgenomic replicons. While wild-type murine fibroblasts are restricted for HCV RNA replication, deletion of IRF-3 alone can facilitate replicon activity in these cells. This effect is thought to be related to the inactivation of the type I interferon synthesis mediated by IRF-3. Additional deletion of IRF-9 to yield IRF-3−/− IRF-9−/− MEFs, which have blocked type I interferon signaling, did not increase HCV replication. Expression of liver-specific miR-122 in MEFs further stimulated the synthesis of HCV replicons in the rodent fibroblasts. The combined effects of miR-122 expression and deletion of IRF-3 produced a cooperative stimulation of HCV subgenome replication. miR-122 and IRF-3 are independent host factors that are capable of influencing HCV replication, and our findings could help to establish mouse models and other cell systems that support HCV growth and particle formation.

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Human immunodeficiency virus facilitates infection/replication of hepatitis C virus in native human macrophages

Blood ◽

10.1182/blood-2003-08-2923 ◽

2004 ◽

Vol 103 (10) ◽

pp. 3854-3859 ◽

Cited By ~ 53

Author(s):

Tomasz Laskus ◽

Marek Radkowski ◽

Joanna Jablonska ◽

Karen Kibler ◽

Jeffrey Wilkinson ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Hiv Infection ◽

Early Stage ◽

Hcv Infection ◽

Hcv Rna ◽

Human Macrophages ◽

Negative Strand ◽

Immunodeficiency Virus

Abstract Hepatitis C virus (HCV) was found to replicate in monocytes/macrophages particularly in patients with human immunodeficiency virus type 1 (HIV-1) infection. This study was undertaken to determine whether HIV facilitates HCV infection of native human macrophages in vitro. Monocytes/macrophages were collected from healthy donors, infected with HIV M-tropic molecular clone, and then exposed to HCV-positive sera. Presence of positive and negative HCV RNA strands was determined with a novel strand-specific quantitative real-time reverse transcription–polymerase chain reaction (RT-PCR). Preceding as well as near-simultaneous infection with HIV made the macrophages more susceptible to infection with HCV; in particular, an HCV RNA–negative strand was detectable almost exclusively in the setting of concomitant HIV infection. Furthermore, HCV RNAload correlated with HIV replication level in the early stage of infection. The ratio of positive to negative strand in macrophages was lower than in control liver samples. HIV infection was also found to facilitate HCV replication in a Daudi B-cell line with engineered CD4 expression. It seems that HIV infection can facilitate replication of HCV in monocytes/macrophages either by rendering cells more susceptible to HCV infection or by increasing HCV replication. This could explain the presence of extrahepatic HCV replication in HIV-coinfected individuals.

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Genotypic Regulation of Type I Interferon Induction Pathways by Frameshift (F) Proteins of Hepatitis C Virus

Journal of Virology ◽

10.1128/jvi.00312-20 ◽

2020 ◽

Vol 94 (15) ◽

Cited By ~ 1

Author(s):

Yi-Ting Lai ◽

Yih-Mei Liou ◽

Fu Hsin ◽

Helene Minyi Liu

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Type I Interferon ◽

Hcv Infection ◽

Type I ◽

Type I Ifn ◽

Coding Sequences ◽

F Protein ◽

Genotype 1A ◽

Core Proteins

ABSTRACT Hepatitis C virus (HCV) has evolved mechanisms to evade innate immunity that are leading to chronic infections. The immunological function of the HCV frameshift (F) protein, which is a frameshift product of core coding sequences, has not been well characterized. The HCV F protein is produced during natural HCV infections and is found most commonly in genotype 1 HCV. In this study, we investigated whether the F protein plays a role in type I interferon (IFN) induction pathways. We engineered F expression constructs from core coding sequences of 4 genotypes (1a, 2a, 3a, and 4a) of HCV as well as the sequences which would only be able to produce core proteins. The peptide lengths and amino acids sequences of F proteins are highly variable. We hypothesized that F proteins from different genotypes might control the type I IFN production and response differently. We found that both IFN-beta (IFN-β) promoter activities are significantly higher in genotype 1a F protein (F1a)-expressing cells. Conversely, the IFN-β promoter activities are lower in genotype 2a F (F2a) protein-expressing cells. We also used real-time PCR to confirm IFN-β mRNA expression levels. By generating chimera F proteins, we discovered that the effects of F proteins were determined by the amino acid sequence 40 to 57 of genotype 1a. The regulation of type I IFN induction pathway is related but not limited to the activity of F1a to interact with proteasome subunits and to disturb the proteasome activity. Further molecular mechanisms of how F proteins from different genotypes of HCV control these pathways differently remain to be investigated. IMPORTANCE Although naturally present in HCV infection patient serum, the virological or immunological functions of the HCV F protein, which is a frameshift product of core coding sequences, remain unclear. Here, we report the effects of the HCV F protein between genotypes and discuss a potential explanation for the differential responses to type I IFN-based therapy among patients infected with different genotypes of HCV. Our study provides one step forward to understanding the host response during HCV infection and new insights for the prediction of the outcome of IFN-based therapy in HCV patients.

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MiR-29c inhibits HCV replicationviaactivation of type I IFN response by targeting STAT3 in JFH-1-infected Huh7 cells

RSC Advances ◽

10.1039/c7ra12815k ◽

2018 ◽

Vol 8 (15) ◽

pp. 8164-8172 ◽

Cited By ~ 1

Author(s):

Yanjing Wang ◽

Yuanyuan Li

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Hcv Infection ◽

Type I ◽

Type I Ifn ◽

Huh7 Cells

MiR-29c, a member of the miR-29 family, has been recognized to play an important role in hepatitis C virus (HCV) infection.

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Hepatitis C virus infection and membranoproliferative glomerulonephritis in Japan.

Journal of the American Society of Nephrology ◽

10.1681/asn.v62220 ◽

1995 ◽

Vol 6 (2) ◽

pp. 220-223

Author(s):

H Yamabe ◽

R J Johnson ◽

D R Gretch ◽

K Fukushi ◽

H Osawa ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Membranoproliferative Glomerulonephritis ◽

Immunoglobulin A ◽

Hcv Infection ◽

Renal Diseases ◽

Hcv Rna ◽

Type I ◽

Hcv Antibody ◽

Polymerase Chain

The prevalence of hepatitis C virus (HCV) infection was determined in 146 adult patients with various types of glomerulonephritis and renal diseases monitored between 1990 and 1993. Serum HCV antibody (HCV Ab) was evaluated, and positive cases were tested for HCV RNA by polymerase chain reaction. HCV infection was present in 1 (1.7%) of 58 cases of immunoglobulin A nephropathy, 0 (0%) of 14 cases of lupus nephritis, 0 (0%) of 12 cases of minimal change nephrosis, and 0 (0%) of 28 cases of other renal diseases, which is similar to the 2% prevalence observed in healthy blood donors in Japan. In contrast, HCV Ab was observed in 2 (8.3%) of 24 cases of membranous nephropathy and 6 (60%) of 10 cases of membranoproliferative glomerulonephritis (MPGN) Type I. The prevalence of HCV infection in MPGN patients was significantly higher than the frequency of HCV infection observed in the other patients with renal diseases (P < 0.001). HCV RNA was present in all cases in which HCV Ab was present. The six patients with HCV-MPGN were similar to the four patients with idiopathic MPGN with respect to age, presence of nephrotic syndrome, and renal dysfunction, but had a higher incidence of liver dysfunction, cryoglobulinemia, rheumatoid factor, and hypocomplementemia (low C3). HCV infection is present in a large percentage of patients with MPGN in Japan and clinically may differ slightly from other cases of MPGN.

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Hepatitis C Virus-Mediated Enhancement of MicroRNA miR-373 Impairs the JAK/STAT Signaling Pathway

Journal of Virology ◽

10.1128/jvi.03085-14 ◽

2015 ◽

Vol 89 (6) ◽

pp. 3356-3365 ◽

Cited By ~ 53

Author(s):

Anupam Mukherjee ◽

Adrian M. Di Bisceglie ◽

Ratna B. Ray

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Signaling Pathway ◽

Human Hepatocytes ◽

Hcv Infection ◽

Janus Kinase ◽

Hcv Rna ◽

Type I ◽

Primary Human Hepatocytes

ABSTRACTHepatitis C virus (HCV) is a serious global health problem and establishes chronic infection in a significant number of infected humans worldwide. Interferon (IFN) and IFN-stimulated genes (ISGs) are amplified during HCV infection but fail to eliminate virus from the liver in a large number of infected patients, and the mechanism is not fully understood. MicroRNAs (miRNAs) have been implicated in the control of many biological processes, including IFN signaling. To gain more insights into the role of cellular miRNAs in possible countermeasures of HCV for suppression of the host antiviral response, a miRNA array was performed by using primary human hepatocytes infected within vitrocell culture-grown HCV. A group of miRNAs were modulated in HCV-infected primary human hepatocytes. We focused on miR-373, as this miRNA was significantly upregulated in HCV-infected primary human hepatocytes. Here, we analyzed the function of miR-373 in the context of HCV infection. HCV infection upregulates miR-373 expression in hepatocytes and HCV-infected liver biopsy specimens. Furthermore, we discovered that miR-373 directly targets Janus kinase 1 (JAK1) and IFN-regulating factor 9 (IRF9), important factors in the IFN signaling pathway. The upregulation of miR-373 by HCV also inhibited STAT1 phosphorylation, which is involved in ISG factor 3 (ISGF3) complex formation and ISG expression. The knockdown of miR-373 in hepatocytes enhanced JAK1 and IRF9 expression and reduced HCV RNA replication. Taken together, our results demonstrated that miR-373 is upregulated during HCV infection and negatively regulated the type I IFN signaling pathway by suppressing JAK1 and IRF9. Our results offer a potential therapeutic approach for antiviral intervention.IMPORTANCEChronic HCV infection is one of the major causes of end-stage liver disease worldwide. Although the recent introduction of direct-acting antiviral (DAA) therapy is extremely encouraging, some infected individuals do not respond to this therapy. Furthermore, these drugs target HCV nonstructural proteins, and with selective pressure, the virus may develop a resistant strain. Therefore, understanding the impairment of IFN signals will help in designing additional therapeutic modalities. In this study, we provide evidence of HCV-mediated upregulation of miR-373 and show that miR-373 impairs IFN signaling by targeting JAK1/IRF9 molecules. The knockdown of miR-373 inhibited HCV replication by upregulating interferon-stimulating gene expression. Together, these results provided new mechanistic insights into the role of miR-373 in HCV infection and suggest a new potential target against HCV infection.

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Prevalence and Risk Factors of Hepatitis C Virus (HCV) in Tehsil Batkhela District Malakand, KPK, Pakistan

International Journal of Contemporary Research and Review ◽

10.15520/ijcrr/2018/9/06/525 ◽

2018 ◽

Vol 9 (06) ◽

pp. 20251-20256

Author(s):

Mudassir Khan ◽

Shahrukh Khan ◽

Shohra Haider ◽

Fazal Jalil ◽

Muhsin Jamal ◽

...

Keyword(s):

Risk Factors ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Skin Color ◽

Significant Risk ◽

Rapid Test ◽

Prevalence Ratio ◽

Hcv Infection ◽

Hcv Rna ◽

Common Symptom

Background: Prevalence of Hepatitis C viral infection and its major risk factors has been found out in population of Batkhela, Khyber Pakhtunkhwa, Pakistan by taking number of volunteers from the interested area. HCV prevalence has not been researched in recent time here in this area, so that’s why we contributed. Materials and Methods: Ab rapid test cassette serum/plasma (USA) kit has been used for the mentioned purpose following by ELISA and finally PCR to find out active infection of virus. ICT positive individuals were reconfirmed by ELISA and then ELISA positive samples were carefully investigated by RT-PCR for Hepatitis C Virus. Results: The study population was of 770 volunteers belonging to the mentioned area of research, 453 males and 317 females. The overall prevalence was found to be 5.32% of HCV in Batkhela. This prevalence ratio was 3.12% in males and 2.20 % in females. 3rd generation ELISA was used to refine ICT positive samples which showed that 37 of the ICT positive samples had antibodies detected by ELISA. To find out active HCV infection, ELISA positive samples were refined by real time PCR which showed 2.98% of prevalence of active HCV infection in Batkhela based on HCV RNA in their blood. Principle Conclusion: Overall prevalence was found 5.32%, contaminated reused syringes and blades at Barbour’s shop, blood transfusion, surgical operations and unhygienic food in stalls etc were found significant risk factors for acquiring HCV infection. Body weakness and pale yellow skin color was common symptom in HCV positive volunteers. Safe sexual activities, blood screening before donation and sterilizing surgical equipment’s can protect us from Hepatitis C Virus.

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Hepatitis C Virus Prevalence and Risk Factors in a Village in Northeastern Romania—A Population-Based Screening—The First Step to Viral Micro-Elimination

Healthcare ◽

10.3390/healthcare9060651 ◽

2021 ◽

Vol 9 (6) ◽

pp. 651

Author(s):

Laura Huiban ◽

Carol Stanciu ◽

Cristina Maria Muzica ◽

Tudor Cuciureanu ◽

Stefan Chiriac ◽

...

Keyword(s):

Risk Factors ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Antiviral Treatment ◽

Population Based ◽

Hcv Infection ◽

World Health ◽

Hcv Rna ◽

Virus Eradication ◽

Tertiary Center

(1) Background: The World Health Organization adopted a strategy for the Global Health Sector on Viral Hepatitis in 2016, with the main objective of eliminating hepatitis C virus (HCV) by 2030. In this work, we aimed to evaluate the prevalence of HCV infection and risk factors in a Romanian village using population-based screening as part of the global C virus eradication program. (2) Methods: We conducted a prospective study from March 2019 to February 2020, based on a strategy as part of a project designed to educate, screen, treat and eliminate HCV infection in all adults in a village located in Northeastern Romania. (3) Results: In total, 3507 subjects were invited to be screened by rapid diagnostic orientation tests (RDOT). Overall, 2945 (84%) subjects were tested, out of whom 78 (2.64%) were found to have positive HCV antibodies and were scheduled for further evaluation in a tertiary center of gastroenterology/hepatology in order to be linked to care. In total, 66 (85%) subjects presented for evaluation and 55 (83%) had detectable HCV RNA. Of these, 54 (98%) completed antiviral treatment and 53 (99%) obtained a sustained virological response. (4) Conclusions: The elimination of hepatitis C worldwide has a higher chance of success if micro-elimination strategies based on mass screening are adopted.

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Prospective study of hepatitis C virus infection in hemodialysis patients by monthly analysis of HCV RNA and antibodies

Canadian Journal of Microbiology ◽

10.1139/w03-065 ◽

2003 ◽

Vol 49 (8) ◽

pp. 503-507 ◽

Cited By ~ 25

Author(s):

Regina Moreira ◽

João Renato Rebello Pinho ◽

Jorge Fares ◽

Isabel Takano Oba ◽

Maria Regina Cardoso ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Hemodialysis Patients ◽

Hcv Infection ◽

Hcv Rna ◽

Hepatitis C Virus Infection ◽

Serum Samples ◽

Hcv Genotypes ◽

High Prevalence ◽

Time Required

The aims of this study were to (i) evaluate the prevalence and the incidence of hepatitis C virus (HCV) infection in hemodialysis patients in two different centers in São Paulo (Brazil), (ii) determine the time required to detect HCV infection among these patients by serology or PCR, (iii) establish the importance of alanine aminotransferase determination as a marker of HCV infection, and (iv) identify the HCV genotypes in this population. Serum samples were collected monthly for 1 year from 281 patients admitted to hospital for hemodialysis. Out of 281 patients, 41 patients (14.6%) were HCV positive; six patients seroconverted during this study (incidence = 3.1/1000 person-month). In 1.8% (5/281) of cases, RNA was detected before the appearance of antibodies (up to 5 months), and in 1.1% (3/281) of cases, RNA was the unique marker of HCV infection. The genotypes found were 1a, 1b, 3a, and 4a. The presence of genotype 4a is noteworthy, since it is a rare genotype in Brazil. These data pointed out the high prevalence and incidence of HCV infection at hemodialysis centers in Brazil and showed that routine PCR is fundamental for improving the detection of HCV carriers among patients undergoing hemodialysis.Key words: HCV genotypes, hemodialysis, hepatitis C, PCR, prevalence, incidence.

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