Coxsackievirus B3 and adenovirus infections of cardiac cells are efficiently inhibited by vector-mediated RNA interference targeting their common receptor

Fechner, Henry; Pinkert, Sandra; Wang, X; Sipo, Isaac; Suckau, Lennart; Kurreck, Jens; Dörner, Andrea; Sollerbrant, Kerstin; Zeichhardt, Heinz; Grunert, H-P; Vetter, Roland; Hp, Schultheiss; Poller, Wolfgang

doi:10.1038/sj.gt.3302948

Cited by 43 publications

(35 citation statements)

References 56 publications

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“…Loading of equal amounts of RNA was verified by rehybridization with a single-stranded antisense b-actin-specific DNA probe as previously described. 49 Hybridized filters were exposed to Kodak BioMax MR film (Sigma-Aldrich).…”

Section: Northern Blot Hybridizationmentioning

confidence: 99%

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

et al. 2010

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Adeno-associated virus (AAV) vectors with capsids of AAV serotype 9 enable an efficient transduction of the heart upon intravenous injection of adult mice but also transduce the liver. The aim of this study was to improve specificity of AAV9 vector-mediated cardiac gene transfer by microRNA (miR)-dependent control of transgene expression. We constructed plasmids and AAV vectors containing target sites (TSs) of liver-specific miR122, miR192 and miR148a in the 3¢ untranslated region (3¢UTR) of a luciferase expression cassette. Luciferase expression was efficiently suppressed in liver cell lines expressing high levels of the corresponding miRs, whereas luciferase expression was unaffected in cardiac myocytes. Intravenous injections of AAV9 vectors bearing three repeats of miR122 TS in the 3¢UTR of an enhanced green fluorescent expression (EGFP) expression cassette resulted in the absence of EGFP expression in the liver of adult mice, whereas the control vectors without miR TS displayed significant hepatic EGFP expression. EGFP expression levels in the heart, however, were comparable between miR122-regulated and control vectors. The liver-specific de-targeting in vivo using miR122 was even more efficient than transcriptional targeting with a cardiac cytomegalovirus (CMV)-enhanced myosin light chain (MLC) promoter. These data indicate that miR-regulated targeting is a powerful new tool to further improve cardiospecificity of AAV9 vectors.

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Section: Northern Blot Hybridizationmentioning

confidence: 99%

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

et al. 2010

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“…CAR knockout resulted in inhibition of CVB3 infections by up to 97% in HL-1 and up to 90% in PNCMs. Adenoviruses were inhibited by only 75% in HL-1, but up to 92% in PNCMs (Fechner et al, 2007). Another host gene, the tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), has been suggested to be a potential target for siRNA to ameliorate CVB3-induced myocarditis.…”

Section: Targeting Host Cellular Genesmentioning

confidence: 99%

“…Thus, adenovirus-derived vector is an ideal carrier for delivery of NA-based antivirals to the heart. This vector has successfully delivered shRNAs targeting the CAR gene in a cardiac-derived HL-1 cell line and isolated PNCMs, resulting in the strong reduction of replication of both CVB3 and adenovirus (Fechner et al, 2007). Lentivirus vectors are derived from HIV.…”

Section: Drug Deliverymentioning

confidence: 99%

Nucleic Acid-Based Strategies for the Treatment of Coxsackievirus-Induced Myocarditis

Yang¹

2011

Myocarditis

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“…polymerases and master regulators) and/or acting early in the virus life cycle are normally selected as the RNAi target(s). However, many RNA viruses such as IBDV can escape the silencing effect due to the high sequence specificity of the RNAi mechanism and rapid mutation rates of viral genes (Fechner et al, 2007). On the other hand, the cellular receptors for virus binding and/or entry are highly conserved proteins and thus can avoid the escape of gene-silencing effects by different viral strains (Haywood, 1994;Fechner et al, 2007).…”

mentioning

confidence: 99%

“…However, many RNA viruses such as IBDV can escape the silencing effect due to the high sequence specificity of the RNAi mechanism and rapid mutation rates of viral genes (Fechner et al, 2007). On the other hand, the cellular receptors for virus binding and/or entry are highly conserved proteins and thus can avoid the escape of gene-silencing effects by different viral strains (Haywood, 1994;Fechner et al, 2007).cHsp90 has been shown to be a functional component of the cellular receptor complex essential for IBDV binding (Lin et al, 2007), which warranted us to investigate the feasibility of inhibiting IBDV infection using an anticHsp90 RNAi strategy. To facilitate effective miRNA selection, we constructed reporter vectors for co-transfection with the miRNA vectors, by which effective miRNAs could be screened easily according to reduction of EGFPpositive cell numbers in the co-transfected cell cultures.…”

mentioning

confidence: 99%

Inhibition of infectious bursal disease virus infection by artificial microRNAs targeting chicken heat-shock protein 90

Yuan

Zhang

Xia

et al. 2012

Journal of General Virology

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Infectious bursal disease virus (IBDV) causes an important disease in young chickens. Chicken heat-shock protein 90 (cHsp90) has been shown to be a functional component of the cellular receptor complex for IBDV infection. This study demonstrates the inhibitory effect of vectorexpressed anti-cHsp90a microRNA (miRNA) on IBDV infection. The reporter vectors pcHsp90a-EGFP and pcHsp90b-EGFP were constructed to facilitate effective miRNA selection. Two anticHsp90a and one anti-cHsp90b miRNA-expression vectors were constructed for a stable transfection study. Poly(A)-tailed RT-PCR detected sequence-specific miRNA transcription in transfected cells. Semiquantitative RT-PCR showed inhibition of cHsp90 transcription in transfected cells. A virus-titration assay showed that the anti-cHsp90a miRNA, but not the anticHsp90b miRNA, had inhibitory effects on IBDV infection. These results suggest that cHsp90a is a functional component of the cellular receptor complex for IBDV infection, and that anti-cHsp90a miRNA could be used as an anti-IBDV reagent.Infectious bursal disease (IBD) is a highly contagious and immunosuppressive disease in young chickens (Sharma et al., 2000). The causative agent, infectious bursal disease virus (IBDV), is a dsRNA virus belonging to the genus Avibirnavirus of the family Birnaviridae (Dobos et al., 1995). IBDV targets the precursors of antibody-producing B-lymphocytes in the bursa of Fabricius (BF), leading to depletion of B-lymphocytes in the BF (Becht, 1980). IBDV infection can cause different mortalities in young chickens, as well as increased susceptibility to other infectious diseases and poor immune response to vaccines (Lukert & Saif, 1997;Van Den Berg, 2000). Despite widely used vaccination programmes, IBD remains one of the major economically important diseases of poultry and thus the development of new strategies for efficient control of the disease is essential (Sharma et al., 2000).RNA interference (RNAi) is a post-transcriptional genesilencing mechanism conserved in eukaryotes ranging from worms to humans (Fire et al., 1998;Meister & Tuschl, 2004). Since its discovery in 1994 as an innate antiviral mechanism, RNAi has become a powerful strategy against a variety of virus infections (Zhou & Rossi, 2011). RNAi has also been used as an in vitro strategy against IBDV, including small interfering RNA (siRNA) against the viral structural protein genes (Gao et al., 2008;Sajjanar et al., 2011) and artificial microRNA (miRNA) targeting VP1 or VP2 transcription (Wang et al., 2009(Wang et al., , 2010. However, IBDV is antigenically variable and undergoing rapid mutation (Müller et al., 2003), which must be addressed in order to establish a viable RNAi approach.IBDV enters the host cell by binding to cellular receptors; several different membrane proteins have been shown to interact with IBDV of different virulence (Delgui et al., 2009; Luo et al., 2010). Lin et al. (2007) demonstrated that chicken heat-shock protein 90 (cHsp90) is a functional component of the cellular receptor complex essential...

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Coxsackievirus B3 and adenovirus infections of cardiac cells are efficiently inhibited by vector-mediated RNA interference targeting their common receptor

Cited by 43 publications

References 56 publications

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

Nucleic Acid-Based Strategies for the Treatment of Coxsackievirus-Induced Myocarditis

Inhibition of infectious bursal disease virus infection by artificial microRNAs targeting chicken heat-shock protein 90

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