Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules

Li, Zhi; He, Ming‐Liang; Yao, Hong; Dong, Qi; Chen, Yangchao; Chan, Cerise Yuen‐Ki; Zheng, Bo‐Jian; Yuen, Kwok‐Yung; Peng, Ying; Sun, Qiang; Yang, Xiao; Lin, Marie Chia‐mi; Sung, Joseph J.Y.; Kung, Hsiang‐Fu

doi:10.5483/bmbrep.2009.42.1.059

Cited by 7 publications

(4 citation statements)

References 32 publications

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“…Hence, HBV is assumed to be susceptible to Rz at both the post-transcription and replication levels. These results are in accordance with those obtained by Li et al [28], who found that a dual-shRNA expression vector (AAV-157i/1694i), which coexpressed 2 shRNAs, targeted the S and X genes of HBV and reduced the levels of HBsAg, HBeAg, and HBV DNA by 87% ± 4%, 80.3% ± 2.6%, and 86.2% ± 7% respectively, at 8 days post-transduction. In a mouse model of prophylactic treatment, HBsAg and HBeAg were reduced to undetectable levels, and the serum HBV DNA level showed at least a 100-fold reduction.…”

Section: Discussionsupporting

confidence: 92%

Efficient inhibition of hepatitis B virus replication by hepatitis delta virus ribozymes delivered by targeting retrovirus

Wang

Pan

et al. 2010

Virol J

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BackgroundHepatitis delta virus (HDV) ribozyme is an attractive molecular tool that can specifically recognize and catalyze the self-cleavage of the viral RNA phosphodiester backbone. However, a major obstacle in the medical application of the HDV ribozyme is the lack of specificity in the delivery of the ribozyme to defined target cells.ResultsThe objective of this study was to determine whether retroviral vectors can deliver the HDV ribozyme into the target cells and to elucidate whether HDV ribozyme plays a role in hepatitis B virus (HBV) replication. In our study, the transduction of helper-free pseudotyped retrovirus, which showed a broad host range, in human hepatoma cells was performed under 2 conditions, that is, in the presence of polymerized human serum albumin (pHSA) and in the absence of pHSA. The transduction ability in the presence of pHSA was higher than in the absence of pHSA. Moreover, HBsAg and HBeAg levels after transductions with pHSA were significantly lower than those in the absence of pHSA, thus indicating that the recombinant retrovirus had HBV-specific cleavage activity and targeted HepG2215 cells.ConclusionsThese data suggest that this system provides a new approach for targeting hepatocytes and has a great potential in gene therapy for HBV infection.

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Section: Discussionsupporting

confidence: 92%

Efficient inhibition of hepatitis B virus replication by hepatitis delta virus ribozymes delivered by targeting retrovirus

Wang

Pan

et al. 2010

Virol J

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“…2) indicated that AAV-shRNA-1 had the greatest antiviral activity, with at least 90% inhibition efficiency at the replication, transcription, or translation level. Since single siRNA administration could be ineffective to inhibit pre-existing mutant HBV genomes 32 , the combination of multiple shRNAs targeting different HBV codon regions was suggested to overcome this problem and has yielded superior antiviral effects 33 . In the present study, the antiviral efficacy of AAV-shRNA-1 + 3 was 2-fold higher than that of a single AAV-shRNA at the DNA and protein levels, whereas a 5-fold higher efficacy was obtained at the RNA level with the use of a dual-shRNA vector.…”

Section: Discussionmentioning

confidence: 99%

Enhanced antiviral and antifibrotic effects of short hairpin RNAs targeting HBV and TGF-β in HBV-persistent mice

Kan

Yan

et al. 2017

Sci Rep

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The hepatitis B virus (HBV) causes acute and chronic liver infection, which may lead to liver cirrhosis and hepatocellular carcinoma. Current treatments including interferons and nucleotide analogs, have limited therapeutic effects, underscoring the need to identify effective therapeutic options to inhibit HBV replication and prevent complications. Previous animal models mimicking chronic HBV infection do not faithfully reflect disease progression in humans. Here, we used our established HBV-persistent mouse line with liver fibrosis to evaluate the efficacy of novel therapies. The combination of two short hairpin RNAs (dual-shRNA) against different coding regions of HBV delivered by a self-complementary AAV vector showed better antiviral effects than single shRNA both in vitro and in HBV-persistent mice. The dual-shRNA also exhibited stronger antifibrotic activity in vivo. Vector carrying shRNA against TGF-β, though did not inhibit HBV replication alone, enhanced the antiviral and antifibrotic activities of single and dual HBV shRNAs. Co-administration of TGF-β shRNA and HBV dual-shRNA decreased HBV DNA, HBV RNA, HBsAg, HBeAg, and liver fibrosis markers in serum and tissues, and improved liver morphology more effectively than single treatments. Our results suggest that the combination of shRNAs against HBV and TGF-β could be developed into a viable treatment for human HBV infection.

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“…The use of an adeno-associated virus (AAV) vector to simultaneously deliver two shRNAs targeting different HBV-related genes (157i targeting the HBV S gene and 1694i targeting the HBV X gene) has been demonstrated to result in greater antiviral effects than vectors that delivered a single shRNA in vitro and in vivo ( 19 ). In the present study, one or two shRNA expression cassettes were subcloned into a pLVX-shRNA1 vector ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Subsequently, the requirement for alternative therapeutic approaches is urgent. Previous studies have suggested that two shRNAs that targeted the HBV S (157i) and X (1694i) genes may be employed to avoid the phenomenon of resistance of HBV mutants to a single siRNA ( 19 ). And the dual shRNA expression vector (AAV-157i-1694i) demonstrated greater antiviral effects in vitro and in vivo compared with those vectors only expressing a single shRNA ( 19 ).…”

Section: Discussionmentioning

confidence: 99%

Therapeutic effects of mesenchymal stem cells combined with short hairpin RNA on liver injury induced by hepatitis B virus infection

Liu

Wang

et al. 2017

Mol Med Report

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The clinical symptoms of chronic hepatitis B virus (HBV) infection include severe liver damage, which is associated with the elimination of the HBV-infected cells by the immune system. It has been suggested that suppression of HBV replication is not sufficient for patients with hepatitis B and the damaged liver function requires restoration. In the present study, mesenchymal stem cells (MSCs) were combined with short hairpin (sh)RNA to treat liver injury and suppress HBV replication in a mouse model. Lx-shRNA157-1694 (an shRNA expression plasmid containing two shRNA expression cassettes) and mouse immortal (mi)MSCs stably expressing shRNA (miMSC-shRNA) were constructed and their suppressive effects on HBV expression were investigated using reverse transcription-polymerase chain reaction (RT-PCR), ELISA and immunofluorescence. Hepatogenic differentiation of miMSC-shRNA was induced in vitro and confirmed by morphology, reverse transcription-semi-quantitative and -quantitative PCR, urea production and Periodic acid-Schiff staining analyses. miMSCs and the shRNA expression plasmid alone or combined with miMSCs stably expressing shRNA were injected into mice. The former therapeutic regimen successfully suppressed HBV expression in sera and liver tissue, whereas the latter only suppressed HBV expression in liver tissue. Analyses of serum alanine aminotransferase levels, aspartate aminotransferase levels, liver weight/body weight ratio percentage and sirius red staining demonstrated marked amelioration of liver injury in mice treated with both therapeutic regimens. The results of the present study suggest that miMSCs combined with shRNA treatment may alleviate liver injury and suppress HBV expression, thus providing a novel potential therapeutic strategy for the treatment of liver injury induced by HBV infection.

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Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules

Cited by 7 publications

References 32 publications

Efficient inhibition of hepatitis B virus replication by hepatitis delta virus ribozymes delivered by targeting retrovirus

Efficient inhibition of hepatitis B virus replication by hepatitis delta virus ribozymes delivered by targeting retrovirus

Enhanced antiviral and antifibrotic effects of short hairpin RNAs targeting HBV and TGF-β in HBV-persistent mice

Therapeutic effects of mesenchymal stem cells combined with short hairpin RNA on liver injury induced by hepatitis B virus infection

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