Iron metabolism and hepatitis C virus (HCV) replication

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Infection with hepatitis C virus (HCV) poses a serious health care problem worldwide and is the leading cause of blood-transmitted chronic hepatitis, liver cirrhosis and hepatocellular cancer. The course of HCV infection is affected by various factors, including the body's iron status. We intend to study the mechanisms by which HCV viral proteins affect cellular iron metabolism in a model cell culture system for subgenomic HCV replication ("replicon"). We have previously utilized this system and demonstrated that administration of iron to replicon cells dramatically decreases HCV replication. The mechanism involves iron-mediated inhibition of the viral RNA polymerase (Fillebeen et al (2005) J. Biol. Chem. 280, 9049-9057). Here we plan to extend these studies and utilize the replicon model system to address how HCV viral proteins affect cellular iron metabolism (specific aim I). On the basis of our findings, the inhibitory effects of interferons in HCV replication and the well-established cross talk between iron metabolism and interferon signaling, we will also explore potential synergistic effects of iron and cytokines in the inhibition of viral replication (specific aim II). Conventional biochemical and molecular biology techniques, such as Western and Northern blotting, will be used to analyze the expression of iron metabolism genes, including ferritin, transferrin receptor, IRP1, IRP2, ferroportin, DMT1 and hepcidin in parent and replicon cells. In addition, high throughput screens, such as microarray analysis and proteomics approaches will be employed to assess how the presence of the replicon interferes with homeostatic responses to iron perturbation and interferon signaling.