Inhibition of adenovirus multiplication by short interfering RNAs directly or indirectly targeting the viral DNA replication machinery

Kneidinger, Doris; Ibrišimović, Mirza; Lion, Thomas; Klein, Reinhard

doi:10.1016/j.antiviral.2012.03.011

Cited by 35 publications

(51 citation statements)

References 62 publications

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“…Alternatively, adenoviral genes other than E1A that more directly link gene expression input to virus replication output can be regulated. In this context, the DNA polymerase or IVa2 genes could be of interest, as their knockdown RNAi proved more effective than that of E1A (37,38). Similarly, aptazyme regulation of the measles virus P or L polymerase genes or of the nucleocapsid N gene may facilitate conditional virus replication, as indicated by RNAi studies (39,40).…”

Section: Discussionmentioning

confidence: 99%

Artificial riboswitches for gene expression and replication control of DNA and RNA viruses

Ketzer

Kaufmann

Engelhardt

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

100

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Aptazymes are small, ligand-dependent self-cleaving ribozymes that function independently of transcription factors and can be customized for induction by various small molecules. Here, we introduce these artificial riboswitches for regulation of DNA and RNA viruses. We hypothesize that they represent universally applicable tools for studying viral gene functions and for applications as a safety switch for oncolytic and live vaccine viruses. Our study shows that the insertion of artificial aptazymes into the adenoviral immediate early gene E1A enables small-molecule-triggered, dosedependent inhibition of gene expression. Aptazyme-mediated shutdown of E1A expression translates into inhibition of adenoviral genome replication, infectious particle production, and cytotoxicity/ oncolysis. These results provide proof of concept for the aptazyme approach for effective control of biological outcomes in eukaryotic systems, specifically in virus infections. Importantly, we also demonstrate aptazyme-dependent regulation of measles virus fusion protein expression, translating into potent reduction of progeny infectivity and virus spread. This not only establishes functionality of aptazymes in fully cytoplasmic genetic systems, but also implicates general feasibility of this strategy for application in viruses with either DNA or RNA genomes. Our study implies that gene regulation by artificial riboswitches may be an appealing alternative to Tet-and other protein-dependent gene regulation systems, based on their small size, RNA-intrinsic mode of action, and flexibility of the inducing molecule. Future applications range from gene analysis in basic research to medicine, for example as a safety switch for new generations of efficiency-enhanced oncolytic viruses.

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Section: Discussionmentioning

confidence: 99%

Artificial riboswitches for gene expression and replication control of DNA and RNA viruses

Ketzer

Kaufmann

Engelhardt

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

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“…Among them, those directed against DNA polymerase, pTP, IVa2, or E1A transcripts have shown the best results, efficiently silencing the respective genes, probably because of the lower amounts of mRNAs transcribed compared with those generated by the strong major late promoters (MLP) of hexon or protease genes [88]. Moreover, given the presence in higher amounts of virus-associated RNAs (VA-RNAs), which seem to counteract RNA interference (RNAi) during late-stage gene expression, the inhibition of HAdV DNA replication might be more beneficial because it would decrease HAdV genome copy numbers and so the copy number of VA-RNA genes [88,95]. In this regard, Kneidinger et al proposed that silencing of the E1A gene, which promotes DNA replication, even when present at low concentrations, could be a potential drug target [88].…”

Section: Adenovirus Dna Replicationmentioning

confidence: 98%

“…Moreover, given the presence in higher amounts of virus-associated RNAs (VA-RNAs), which seem to counteract RNA interference (RNAi) during late-stage gene expression, the inhibition of HAdV DNA replication might be more beneficial because it would decrease HAdV genome copy numbers and so the copy number of VA-RNA genes [88,95]. In this regard, Kneidinger et al proposed that silencing of the E1A gene, which promotes DNA replication, even when present at low concentrations, could be a potential drug target [88].…”

Section: Adenovirus Dna Replicationmentioning

confidence: 99%

“…The E1A protein has an important role in the regulation of host gene expression and is also necessary for the expression of the early genes E1B, E2, E3 and E4 from different promoters as well as for HAdV DNA replication [89,91,92]. Moreover, for HAdV DNA replication to occur requires three essential proteins: the viral DNA polymerase, the preterminal protein (pTP), and the DNA-binding protein, all of which are potential targets for new antiHAdV drugs [88]. Among the different potential targets of HAdV, the DNA replication process (Figure 3) has been the most explored, and several molecules other than cidofovir have been described [7,19,50].…”

Section: Adenovirus Dna Replicationmentioning

confidence: 99%

“…Before HAdV DNA replication, the transcription of adenovirus early region 1A (E1A) by cellular RNA polymerase II occurs from the E1A promoter [87][88][89][90]. The E1A protein has an important role in the regulation of host gene expression and is also necessary for the expression of the early genes E1B, E2, E3 and E4 from different promoters as well as for HAdV DNA replication [89,91,92].…”

Section: Adenovirus Dna Replicationmentioning

confidence: 99%

See 2 more Smart Citations

Antiadenovirus drug discovery: potential targets and evaluation methodologies

Martínez-Aguado

Serna-Gallego

Marrugal-Lorenzo

et al. 2015

Drug Discovery Today

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Adenovirus persistence, reactivation, and clinical management

2019

Self Cite

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Adenoviral infections continue posing a major threat in severely immunocompromised patients including particularly allogeneic stem cell transplant recipients. Although exogenous infections occur in some instances, the majority of invasive events appear to arise from viral reactivation. In the pediatric setting, adenoviruses were demonstrated to persist in the gastrointestinal tract, and the intestinal epithelium serves as the main site of viral replication preceding invasive infection. Regular monitoring of serial stool samples for the presence and load of adenoviruses has therefore become a routine diagnostic tool for post‐transplant patient surveillance, and can serve as a trigger for early initiation of treatment. In the adult setting, the source of infection or reactivation is less clear, and monitoring of peripheral blood specimens is the predominant approach for patient surveillance. Timely initiation of antiviral treatment is reportedly required for prevention or successful control of disseminated disease mediated by adenoviruses, and appropriate diagnostic monitoring is therefore of paramount importance. Currently available antiviral agents and immune therapeutic approaches have not been able to entirely overcome the life‐threatening courses of invasive adenoviral infections in the immunocompromised clinical setting.

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Inhibition of adenovirus multiplication by short interfering RNAs directly or indirectly targeting the viral DNA replication machinery

Abstract: Highlights ► SiRNAs inhibit adenovirus multiplication. ► Adenoviral DNA replication is a key target for siRNA-mediated inhibition. ► SiRNAs can delay the death of adenovirus-infected cells.

Cited by 35 publications

References 62 publications

Artificial riboswitches for gene expression and replication control of DNA and RNA viruses

Artificial riboswitches for gene expression and replication control of DNA and RNA viruses

Antiadenovirus drug discovery: potential targets and evaluation methodologies

Adenovirus persistence, reactivation, and clinical management

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