Geteikende inhibisie van Hepatitis-B-virus geenuitdrukking

Abstract

Inhibition of Rous Sarcoma Virus replication in cell culture with oligonucleotides complementary to strategic regions of the viral genome first suggested the therapeutic potential of antisense technology. Complementary oligonucleotides were subsequently employed to inhibit Human Immunodeficiency Virus, Vesicular Stomatitis Virus, Influenza Virus, Herpes Simplex Virus, and the expression of various oncogenes. Because infection with Hepatitis B Virus (HBV) is endemic throughout much of the world, possibly affecting 200 million people, and is associated with serious clinical consequences such as liver cirrhosis and hepatocellular carcinoma, we investigated the possibility of inhibiting HBV gene expression by anti sense technology. Anti sense 9-, 1 5-, and 21-mer oligonucleotides were targeted against two conserved regions in the HBsAg ORF, one overlapping the initiation codon of the S gene and the other 300 bp downstream. HBsAg expression by PLC/PRF/5 hepatocellular carcinoma cells was determined after transfection with the various antisense oligonucleotides. Pronounced inhibition of HBsAg expression was observed; a single dose of 20 µM of a 15-mer targeted at the initiation region of the gene resulted in more than 60% inhibition of HBsAg expression over a period of five days. Our results strongly suggested that antisense technology may be employed to inhibit HBV gene expression.