Vivo-morpholino oligomers strongly inhibit dengue virus replication and production

Abstract

Dengue virus (DENV) infection is a worldwide public health problem, which can cause severe dengue hemorrhagic fever (DHF) and life-threatening dengue shock syndrome (DSS). There are currently no anti-DENV drugs available, and there has been an intensive search for effective anti-DENV agents that can inhibit all four DENV serotypes. In this study, we tested whether vivo-morpholino oligomers (vivo-MOs), whose effect on DENV infection has not previously been studied, can inhibit DENV infection. Vivo-MOs were designed to target the top of 3’ stem-loop (3’ SL) in the 3’ UTR of the DENV genome and tested for inhibition of DENV infection in monkey kidney epithelial (Vero) cells and human lung epithelial carcinoma (A549) cells. The results showed that vivo-MOs could bind to a DENV RNA sequence and markedly reduce DENV-RNA, protein, and virus production in infected Vero and A549 cells. Vivo-MOs at a concentration of 4 µM could inhibit DENV production by more than 104-fold when compared to that of an untreated control. In addition, vivo-MOs also inhibited DENV production in U937 cells and primary human monocytes. Therefore, vivo-MOs targeting to the 3’ SL in the 3’ UTR of DENV genomes are effective and have the potential to be developed as anti-DENV agents.

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Acknowledgements

This work was supported by a Mahidol University Grant (R15610004) and the Thailand Research Fund (TRF IRG 5980006). PP was supported by a TRF-Royal Golden Jubilee (RGJ) Ph.D. Scholarship (PHD/0367/2552). PYo was supported by Siriraj Graduate and Thesis Scholarships. MJ and PY were supported by Chaloemprakiat Grants, Faculty of Medicine Siriraj Hospital, Mahidol University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Correspondence to Pa-thai Yenchitsomanus.

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The use of human peripheral blood mononuclear cells (PBMCs) from healthy donors was ethically approved by the Siriraj Institutional Review Board, Faculty of Medicine Siriraj Hospital, Mahidol University.

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Supplementary Figure 1 Binding of vivo-MO-2 to biotin-tagged DENV-2 RNA. Vivo-MO-2 at a concentration 50, 100, or 200 times that of the RNA target was added to 5’-end-biotin-tagged DENV-2 RNA (2 nM). Vivo-MO-NC (negative control) was added at a concentration 200 times that of the RNA target. The binding of vivo-MO-2 to biotin-tagged DENV-2 RNA was detected by non-denaturing gel electrophoresis. The data shown are the results from two experiments. (TIFF 414 kb)

Supplementary Figure 2 Effect of vivo-MO-1 on DENV-2 in A549 cells. A549 cells were infected with DENV-2 for 2 hours and treated with vivo-MO-NC or vivo-MO-1 at a concentration of 0.5, 1, 2, 4, 6, or 8 μM. The cells were incubated for 24 hours after infection. (A) Cell viability measured by PlestoBLUETM Cell Viability assay. (B) Virus production in the supernatants, measured by focus assay. (C) Intracellular DENV-2 E antigen detected by immunofluorescence assay (IFA). (TIFF 959 kb)

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Phumesin, P., Junking, M., Panya, A. et al. Vivo-morpholino oligomers strongly inhibit dengue virus replication and production. Arch Virol 163, 867–876 (2018). https://doi.org/10.1007/s00705-017-3666-9

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