Development of hepatitis C virus suicide therapy employing the viral protease

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Hepatitis C virus (HCV) inactivates the innate immunity pathway, which leads from retinoic acid-inducible gene (RIG)-I to IFNss-induction, by the specific digestion of IFN promoter-stimulator (IPS)-1 on the mitochondrial outer membrane, by the action of viral protease NS3/4a. This results in the malfunction of IFN-activation system in the liver, which is considered to play a central role in the persistence of HCV infection and the resistance to IFN treatment. We have attempted to establish “HCV suicide thrapy” in that sending the transcription module (HCV-protease activated module; CPAM, inactivation form) into the liver by the utilization of NS3/4a protease. It is supposed that inactivated form of the CPAM is activated by the action of NS3/4a protease in HCV-infected cells, leading to the activation of intrahepatic innate immunity and complete eradication of HCV from the liver.In 2011, we constructed the CPAM module that carries IRF7 in conjunction with mitochondrial transfer signal and NS3/4a protease digestion motif (c503 amino acid residues), which derived from IPS-1.In 2012, The CPAM was introduced into Huh-7 cells in which the HCV subgenomic replicon replicates and NS3/4a protease is produced. IRF7 was actually transferred from the cytoplasm to nuclei. In addition, we constructed the recombinant adenovirus that carries the CPAM. This was introduced into Huh-7 cells that support the replication of HCV JFH-1 strain to evaluate the load-reducing effect of CPAM. The JFH-1 loads weresignificantly reduced in CPAN- introduced Huh-7 cells compared to control Huh-7 cells, demonstrating the proof of concept of this HCV suicide therapy using NS3/4a protease.