The sustained antiviral response of chronic HCV infection has been improved recently by the use of the protease inhibitor (telaprevir or boceprevir) along with interferon alpha (IFN-?) and ribavirin. However, results of recent clinical trials indicate that the final outcome of triple combination therapy is strongly dependent on the patient's initial response to IFN-? plus ribavirin and the host genetic variation of IL-28 gene polymorphism. The biological mechanisms underpinning this association remain unknown. The current application explores mechanisms for how the IFN-? axis is linked to the success of IFN-?/ribavirin treatment using a stable and persistently infected HCV cell culture system. The proposal is based on several exciting and encouraging preliminary studies. We have developed a persistently HCV infected cell culture system that supports high- level viral replication to address the antiviral mechanisms of IFN-? and ribavirin combination treatment. We reported that HCV infection itself induces ER-stress and autophagy response that selectively down regulates the type-I IFN receptor but not the type-II or type-III IFN receptor, impaired Stat1/Stat phopshorylation, nuclear translocation, defective Jak-Stat signaling and impaired antiviral response. We have reported that ribavirin and IFN? each inhibit HCV IRES translation and synergistically inhibit HCV replication. The ribavirin resistance mechanism relates to the impaired ribavirin uptake due to the down-regulated expression nucleoside transporters in HCV cell culture. Our results indicate that IFN-? has a strong antiviral action against HCV and it clears HCV replication to a completion in Jak-Stat defective and persistently infected HCV cell culture system through activation of Stat3, AKT and MAPK pathways suggesting that the type III IFN system plays a very important role in the clearance of HCV infection. In this R01 application, we will focus on understanding the mechanism underlying the association between IL28B polymorphisms and IFN-?/ribavirin antiviral mechanisms of HCV infection in a persistent infectious cell culture system in hepatocyte cell lines with a different genetic background of IL-28B gene. Our hypothesis is that hepatitis C virus infection itself creates defective Jak-Stat signaling by down regulating the expression of IFNAR1 that leads to impaired response to pegylated IFN-? and ribavirin. To test this hypothesis we propose the following three Specific Aims.
Aim 1 : To test hypothesis that the IFN-? axis is important for HCV clearance and treatment using a persistently infected HCV cell culture.
Aim 2 : Study the mechanisms by which IFN-? overcome IFN-? resistance of persistent HCV infection.
Aim 3 : To investigate the synergy and resistance mechanism of ribavirin in HCV infection. Since IFN-? clears HCV replication in Jak-Stat defective IFN-? resistant cells, understanding the signal transduction and anti-viral mechanism of IFN-? proposed in this grant application should open novel strategies for the treatment of chronic HCV infection.

Public Health Relevance

Hepatitis C virus infection is the leading cause of chronic liver disease, liver cirrhosis and hepatocellular carcinoma in the United States. Interferon alpha plus ribavirin along with protease inhibitor is the FDA approved care for chronic HCV infection. Recently, genome-wide association studies have identified that the certain host IL-28B gene polymorphisms is critical for patient's response to antiviral drug treatment and spontaneous clearance. The mechanism underpinning this association is not clear. This grant application seeks to explore the genetic association of IL-28B with interferon/ribavirin treatment clearance of chronic HCV infection in tissue culture. The knowledge gained from these investigations will increase our understanding of hepatitis C virus resistance mechanisms to interferon alpha /ribavirin and explore the therapeutic utility of a new type-III interferon as an alternative stratgy to overcome resistance, clear the virus infection and prevent liver cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI103106-02
Application #
8706035
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Koshy, Rajen
Project Start
2013-08-01
Project End
2017-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Tulane University
Department
Pathology
Type
Schools of Medicine
DUNS #
City
New Orleans
State
LA
Country
United States
Zip Code
70118
Ferraris, Pauline; Chandra, Partha K; Panigrahi, Rajesh et al. (2016) Cellular Mechanism for Impaired Hepatitis C Virus Clearance by Interferon Associated with IFNL3 Gene Polymorphisms Relates to Intrahepatic Interferon-λ Expression. Am J Pathol 186:938-51
Dash, Srikanta; Chava, Srinivas; Aydin, Yucel et al. (2016) Hepatitis C Virus Infection Induces Autophagy as a Prosurvival Mechanism to Alleviate Hepatic ER-Stress Response. Viruses 8:
Aboulnasr, Fatma; Hazari, Sidhartha; Nayak, Satyam et al. (2015) IFN-λ Inhibits MiR-122 Transcription through a Stat3-HNF4α Inflammatory Feedback Loop in an IFN-α Resistant HCV Cell Culture System. PLoS One 10:e0141655
Panigrahi, Rajesh; Chandra, Partha K; Ferraris, Pauline et al. (2015) Persistent hepatitis C virus infection impairs ribavirin antiviral activity through clathrin-mediated trafficking of equilibrative nucleoside transporter 1. J Virol 89:626-42
Peralta, Donna V; Heidari, Zahra; Dash, Srikanta et al. (2015) Hybrid paclitaxel and gold nanorod-loaded human serum albumin nanoparticles for simultaneous chemotherapeutic and photothermal therapy on 4T1 breast cancer cells. ACS Appl Mater Interfaces 7:7101-11
Kurt, Ramazan; Chandra, Partha K; Aboulnasr, Fatma et al. (2015) Chaperone-Mediated Autophagy Targets IFNAR1 for Lysosomal Degradation in Free Fatty Acid Treated HCV Cell Culture. PLoS One 10:e0125962
Bao, Lili; Chandra, Partha K; Moroz, Krzysztof et al. (2014) Impaired autophagy response in human hepatocellular carcinoma. Exp Mol Pathol 96:149-54
Gunduz, Feyza; Mallikarjun, Chaithanya; Balart, Luis A et al. (2014) Interferon alpha induced intrahepatic pSTAT1 inversely correlate with serum HCV RNA levels in chronic HCV infection. Exp Mol Pathol 96:36-41
Chandra, Partha K; Gunduz, Feyza; Hazari, Sidhartha et al. (2014) Impaired expression of type I and type II interferon receptors in HCV-associated chronic liver disease and liver cirrhosis. PLoS One 9:e108616
Chandra, Partha K; Bao, Lili; Song, Kyoungsub et al. (2014) HCV infection selectively impairs type I but not type III IFN signaling. Am J Pathol 184:214-29

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