A phosphorothioate antisense oligodeoxynucleotide specifically inhibits coxsackievirus B3 replication in cardiomyocytes and mouse hearts

Abstract

Antisense oligodeoxynucleotides (AS-ODNs) are promising therapeutic agents for the treatment of virusinduced diseases. We previously reported that coxsackievirus B3 (CVB3) infectivity could be inhibited effectively in HeLa cells by phosphorothioate AS-ODNs complementary to different regions of the 5 0 and 3 0 untranslated regions of CVB3 RNA. The most effective target is the proximal terminus of the 3 0 untranslated region. To further investigate the potential antiviral role of the AS-ODN targeting this site in cardiomyocytes (HL-1 cell line), corresponding AS-ODN (AS-7) was transfected into the HL-1 cells and followed by CVB3 infection. Analyses by RT-PCR, Western blotting and plaque assay demonstrated that AS-7 strongly inhibits viral RNA and viral protein synthesis as compared to scrambled AS-ODNs. The percent inhibitions of viral RNA transcription and capsid protein VP1 synthesis were 87.6 and 40.1, respectively. Moreover, AS-7 could inhibit ongoing CVB3 infection when it was given after virus infection. The antiviral activity was further evaluated in a CVB3 myocarditis mouse model. Adolescent A/J mice were intravenously administrated with AS-7 or scrambled AS-ODNs prior to and after CVB3 infection. Following a 4-day therapy, the myocardium CVB3 RNA replication decreased by 68% and the viral titers decreased by 0.5 log 10 in the AS-7-treated group as compared to the group treated with the scrambled AS-ODNs as determined by RT-PCR, in situ hybridization and viral plaque assay. Taken together, our results demonstrated a great potential for AS-7 to be further developed into an effective treatment towards viral myocarditis as well as other diseases caused by CVB3 infection.

Pairwise network mechanisms in the host signaling response to coxsackievirus B3 infection

“…To investigate the host network response to CVB3 in a uniform cell population, we used HL-1 murine cardiomyocytes (17). These cells can be infected with CVB3, support viral replication, and undergo virus-induced cell death (18).…”

Inhibition of glycogen synthase kinase 3β suppresses coxsackievirus-induced cytopathic effect and apoptosis via stabilization of β-catenin

“…48 Cell supernatant was serially diluted and overlaid on monolayer of HeLa cells. Following 1 h of incubation, medium was removed and complete DMEM containing 0.75% agar was overlaid.…”

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

“…Wang et al, 2001). The importance of these sites for ASON binding was further confirmed by in vivo evaluation using a murine myocarditis model, although the antiviral efficiency is not as high as that obtained from in vitro evaluation (Yuan et al, 2004). To improve the stability of the oligomers, our group designed eight phosphorodiamidate morpholino oligomers (PMO) targeting both the sense and antisense strands of the CVB3 replication intermediate.…”