Inhibition of hepatitis C virus gene expression by adenoviral vectors encoding antisense RNA in vitro and in vivo

Gonzalez‐Carmona, Maria A.; Vogt, Annabelle; Heinicke, Thomas; Quasdorff, Maria; Hoffmann, Per; Yildiz, Yildiz; Schneider, Cindy; Serwe, Matthias; Bartenschlager, Ralf; Sauerbruch, Tilman; Caselmann, Wolfgang H.

doi:10.1016/j.jhep.2010.11.010

Cited by 10 publications

(9 citation statements)

References 32 publications

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“…There have been numerous reports that RNAi-based agents such as PF-05095808 deliver antiviral activity in HCV replicon systems (14,31,35,45). Our studies were intended to go beyond this proof of principle and to identify methodologies that would enable the linking of in vitro activity with the potential to predict in vivo efficacy.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Characterization of the Activity of PF-05095808, a Novel Biological Agent for Hepatitis C Virus Therapy

Lavender¹,

Brady²,

Burden³

et al. 2012

Antimicrob Agents Chemother

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PF-05095808 is a novel biological agent for chronic hepatitis C virus (HCV) therapy. It comprises a recombinant adenoassociated virus (AAV) DNA vector packaged into an AAV serotype 8 capsid. The vector directs expression of three short hairpin RNAs (shRNAs) targeted to conserved regions of the HCV genome. These shRNAs are processed by the host cell into the small interfering RNAs which mediate sequence-specific cleavage of target regions. For small-molecule inhibitors the key screens needed to assess in vitro activity are well defined; we developed new assays to assess this RNA interference agent and so to understand its therapeutic potential. Following administration of PF-05095808 or corresponding synthetic shRNAs, sequence-specific antiviral activity was observed in HCV replicon and infectious virus systems. To quantify the numbers of shRNA molecules required for antiviral activity in vitro and potentially also in vivo, a universal quantitative PCR (qPCR) assay was developed. The number of shRNA molecules needed to drive antiviral activity proved to be independent of the vector delivery system used for PF-05095808 administration. The emergence of resistant variants at the target site of one shRNA was characterized. A novel RNA cleavage assay was developed to confirm the spectrum of activity of PF-05095808 against common HCV clinical isolates. In summary, our data both support antiviral activity consistent with an RNA interference mechanism and demonstrate the potential of PF-05095808 as a therapeutic agent for chronic HCV infection.

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Section: Discussionmentioning

confidence: 99%

In Vitro Characterization of the Activity of PF-05095808, a Novel Biological Agent for Hepatitis C Virus Therapy

Lavender¹,

Brady²,

Burden³

et al. 2012

Antimicrob Agents Chemother

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“…The SVR rate for G4 HCV infected patients after Peg-IFN/RBV treatment is approximately 60% [16]. Currently, novel compounds against the HCV-NS3 protease or the HCV-NS5B RNA-polymerase have entered clinical trials, showing high antiviral potency [17]. However, rapid HCV drug resistance to these agents has been shown to limit their efficacy, necessitating a combination with PEG-IFN and ribavirin, which may cause a wide range of serious adverse reactions [18, 19].…”

Section: Introductionmentioning

confidence: 99%

In VitroInhibition of Hepatitis C Virus by Antisense Oligonucleotides in PBMC Compared to Hepatoma Cells

Youssef

Fahmy

Omran

et al. 2014

BioMed Research International

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Aim. To assess the efficiency of phosphorothioate antisense oligodeoxynucleotide 1 (S-ODN1) on HCV translation inhibition in PBMC compared to hepatoma cells in vitro for the first time. Materials and Methods. The study included 34 treatment naive HCV patients. IRES domain III and IV sequence variations were tested in 45 clones from 9 HCV patients. PBMC of HCV positive patients were subjected to S-ODN in vitro. Concomitantly HepG2 cells infected by the same patient's serum were also treated with S-ODN1 for 24 and 48 hours. Cellular RNA was tested for HCV plus and minus strands by reverse transcription polymerase chain reaction (RT-PCR). Results. Sequence variations were seen in HCV IRES domain III only while domain IV was conserved among all the tested patient's clones. S-ODN1 successfully inhibited HCV translation in HepG2 cells, while in PBMC inhibition was partial. Conclusion. HCV IRES domain IV is more conserved than domain IIId in genotype 4 HCV patients. S-ODN against HCV IRES domain IV was not efficient to inhibit HCV translation in PBMC under the study conditions. Further studies testing other S-ODN targeting other HCV IRES domains in PBMC should be done.

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“…Target RNA cleavage or translation inhibition 5'-UTR and 3'-UTR [93][94][95][96][100][101][102] Protein coding regions [103][104][105][106][107][108][109] Antisense oligonucleotide Bind to complementary RNAs and suppress the access to cellular machinery, thereby inhibiting expression or function of the targeted RNAs 5'-UTR [134][135][136][137] Completion of Phase Ⅱ…”

Section: Rnaimentioning

confidence: 99%

“…McCaffrey et al [136] demonstrated that morpholino phosphoramidate antisense oligonucleotides (morpholinos) complementary to the HCV 5'-UTR specifically inhibited HCV IRES-dependent luciferase translation by up to 95% for at least 6 d in mouse liver. Moreover, an adenoviral vector-expressing an RNA ASO has been reported to block HCV replication in the HCV replicon and in the infectious HCV JFH-1 cell culture system by up to 40% and 76%, respectively [137] . Recently, a very promising ASO against HCV was reported with LNA-modified Miravirsen (SPC3649; Santaris Pharmaceuticals), which is directed against microRNA 122 (miR-122) [57] .…”

Section: Antisense Oligonucleotidementioning

confidence: 99%

Prospects for nucleic acid-based therapeutics against hepatitis C virus

Lee

Kim

Lee

2013

WJG

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In this review, we discuss recent advances in nucleic acid-based therapeutic technologies that target hepatitis C virus (HCV) infection. Because the HCV genome is present exclusively in RNA form during replication, various nucleic acid-based therapeutic approaches targeting the HCV genome, such as ribozymes, aptamers, siRNAs, and antisense oligonucleotides, have been suggested as potential tools against HCV. Nucleic acids are potentially immunogenic and typically require a delivery tool to be utilized as therapeutics. These limitations have hampered the clinical development of nucleic acid-based therapeutics. However, despite these limitations, nucleic acid-based therapeutics has clinical value due to their great specificity, easy and large-scale synthesis with chemical methods, and pharmaceutical flexibility. Moreover, nucleic acid therapeutics are expected to broaden the range of targetable molecules essential for the HCV replication cycle, and therefore they may prove to be more effective than existing therapeutics, such as interferon-α and ribavirin combination therapy. This review focuses on the current status and future prospects of ribozymes, aptamers, siRNAs, and antisense oligonucleotides as therapeutic reagents against HCV.

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Inhibition of hepatitis C virus gene expression by adenoviral vectors encoding antisense RNA in vitro and in vivo

Cited by 10 publications

References 32 publications

In Vitro Characterization of the Activity of PF-05095808, a Novel Biological Agent for Hepatitis C Virus Therapy

In Vitro Characterization of the Activity of PF-05095808, a Novel Biological Agent for Hepatitis C Virus Therapy

In VitroInhibition of Hepatitis C Virus by Antisense Oligonucleotides in PBMC Compared to Hepatoma Cells

Prospects for nucleic acid-based therapeutics against hepatitis C virus

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