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Exploring gene-deleted adenoviral vectors for delivery of short hairpin RNAs and reduction of hepatitis B virus infection in mice

✍ Scribed by Christina Rauschhuber; Hui Xu; Felix H. Salazar; Patricia L. Marion; Anja Ehrhardt


Book ID
102890999
Publisher
John Wiley and Sons
Year
2008
Tongue
English
Weight
706 KB
Volume
10
Category
Article
ISSN
1099-498X

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✦ Synopsis


Abstract

Background

RNA interference based therapeutic approaches hold promise for the treatment of patients chronically infected with hepatitis B virus (HBV). To conquer HBV infection, long‐term suppression of target transcripts in all hepatocytes without toxic effects may be required. The present study explored gene‐deleted adenoviral vectors (GD‐AdV) lacking all viral coding sequences for delivery of the previously described short hairpin RNA (shRNA) HBVU6no.2, which was demonstrated to result in post‐transcriptional knock‐down of HBV transcripts.

Methods

We established conditions for shRNA delivery expressed from GD‐AdV in vitro and in vivo and observed up to 96% shRNA‐mediated knockdown of luciferase expressed in mouse liver. To investigate in vivo efficacy of HBVU6no.2 expressed from a GD‐AdV, we explored a transient and a transgenic mouse model for HBV infection.

Results

We observed an up to 68% drop in serum HBV surface antigen (HBsAg) levels in the transient and the transgenic mouse model for HBV infection, respectively. Interestingly, we detected an up to 86% drop in HBsAg levels in both animal models after administration of a control GD‐AdV encoding β‐galactosidase. In concordance with reduced serum HBsAg levels, we observed reduced HBV replication as demonstrated by Southern blot analysis of HBV genomes.

Conclusions

The present study demonstrates that GD‐AdV can be used against HBV infection but the design of DNA sequences including shRNAs contained in the vector and virus–host interactions during superinfection needs to be carefully considered. Copyright © 2008 John Wiley & Sons, Ltd.