Antisense RNA Complementary to Hepatitis B Virus Specifically Inhibits Viral Replication

GASTROENTEROLOGY 1998;115:702-713

JASPERZU PUTLITZ*, STEFAN WIELAND, HUBERT E. BLUM, and JACK R. WANDS*

· Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center and Harvard Medical School, Boston, Massachusetts; and Department of Internal Medicine II, University of Freiburg, Freiburg, Germany

Background & Aims:

Chronic infection with the Hepatitis B virus (HBV) is a major public health problem, and currently available therapies have limited efficacy. Gene therapy strategies for HBV infection are under active investigation. We evaluated the potential of antisense RNA transcribed from antisense genes to interfere with HBV replication.

Methods:

Subgenomic fragments of the HBV genome were studied with respect to the property of inhibiting HBV replication when intracellularly expressed in the antisense orientation. Results: Antisense RNAs derived from the HBV genome specifically inhibited HBV replication and antigen expression in human hepatocellular carcinoma cells by 60%-75%. DNA sequences corresponding to the identified RNAs had no effect on HBV replication, indicating that inhibitory effects are mediated by RNA. Transcripts corresponding to the inhibitory subgenomic fragments were present at high levels. One antisense RNA was found to reduce the amount of pregenomic RNA encapsidated into core particles as a molecular mechanism of antiviral effects.

Conclusions:

Certain antisense RNA molecules will have substantial antiviral effects against HBV. Antisense RNAs derived from the HBV genome are promising candidates as antiviral agents and may serve as novel tools to identify functionally important regions of HBV transcripts.

GASTROENTEROLOGY 1998;115:702-713

© 1998 by The American Gastroenterological Association