Primary research

Antisense oligonucleotide inhibition of hepatitis C virus genotype 4 replication in HepG2 cells

Mostafa K El Awady¹ , Noha G Badr El Din¹ , Wael T El Garf¹ , Samar S Youssef¹ , Moataza H Omran¹ , Jasmin El Abd¹ and Said A Goueli^2,3

¹  Department of Biomedical Technology, National Research Center, Dokki

²  Research and Development, Promega Corp., University of Wisconsin, Madison, USA

³  Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, USA

author email corresponding author email

Cancer Cell International 2006, 6:18doi:10.1186/1475-2867-6-18

Published:	27 June 2006

Abstract

Background

Hepatitis C (HCV) viral infection is a serious medical problem in Egypt and it has a devastating impact on the Egyptian economy. It is estimated that over 15% of Egyptians are infected by the virus and thus finding a cure for this disease is of utmost importance. Current therapies for hepatitis C virus (HCV) genotype 4 with interferon/ribavirin have not been successful and thus the development of alternative therapy for this genotype is disparately needed.

Results

Although previous studies utilizing viral subgenomic or full cDNA fragments linked to reporter genes transfected into adhered cells or in a cell free system showed promise, demonstration of efficient viral replication was lacking. Thus, we utilized HepG2 cells infected with native HCV RNA genomes in a replication competent system and used antisense phosphorothioate Oligonucleotides (S-ODN) against stem loop IIId and the AUG translation start site of the viral polyprotein precursor to monitor viral replication. We were able to show complete arrest of intracellular replication of HCV-4 at 1 uM S-ODN, thus providing a proof of concept for the potential antiviral activity of S-ODN on native genomic replication of HCV genotype 4.

Conclusion

We have successfully demonstrated that by using two S-ODNs [(S-ODN1 (nt 326–348) and S-ODN-2 (nt 264–282)], we were able to completely inhibit viral replication in culture, thus confirming earlier reports on subgenomic constructs and suggesting a potential therapeutic value in HCV type 4.