Antiviral activity of modified oligonucleotides in human lymphoid cells infected with a strain of HIV‐1
https://doi.org/10.18470/1992-1098-2024-4-5
Abstract
Aim. To investigate the cell entry and antiretroviral properties of modified oligodeoxyribonucleotides targeting highly conserved regions of the HIV‐1 genome: the study was conducted using modified oligonucleotides containing phosphorothioate, phosphorylguanidine or dodecyl moieties.
The ability of oligonucleotides to inhibit the virus was studied using a model of lymphoid culture of human MT4 cells infected with a characterised high‐productive strain of HIV‐1 belonging to the subtype A6 genovariant widely spread in the Russian Federation. The entry of oligonucleotides into MT4 cells was assessed by confocal microscopy.
It was shown that phosphorothioate‐ and dodecyl‐containing oligonucleotides are able to penetrate inside cells without the use of any additional transfection agents. It was found that phosphorothioate oligonucleotides transfected in duplex with dodecyl‐containing derivatives were localized inside the cell in the nucleus and nucleolus and were able to inhibit the reproduction of a highly productive HIV‐1 strain. A 50% toxic dose (TC50) and a 50% inhibitory concentration (IC50) were determined for the oligonucleotide derivatives studied, the value of which was less than 0,5 μM in the case of phosphorothioate oligonucleotides.
The data obtained indicate the ability of the modified oligonucleotides studied to be used as potential antiviral agents against HIV‐1.
Keywords
HIV,
modified oligodeoxyribonucleotides,
oligonucleotides delivery,
antiretroviral activity of modified oligonucleotides
Supporting agencies: State Assignment no. 1/23 (FBRI SRC VB “Vector” Rospotrebnadzor) supported this research.
About the Authors
L. G. Gotfrid
Vector State Research Centre of Virology and Biotechnology
Russian Federation
Russian Federation
Ludmila G. Gotfrid, Junior Researcher
ABK 12A, Koltsovo, Novosibirsk Oblast, 630559. Tel. +79134506795
A. S. Pavlova
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
Russian Federation
Russian Federation
Anna S. Pavlova
Novosibirsk
M. S. Kupryushkin
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
Russian Federation
Russian Federation
Maxim S. Kupryushkin
Novosibirsk
I. A. Pyshnaya
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
Russian Federation
Russian Federation
Inna A. Pyshnaya
Novosibirsk
N. M. Gashnikova
Vector State Research Centre of Virology and Biotechnology, Rospotrebnadzor
Russian Federation
Russian Federation
Natalya M. Gashnikova
Koltsovo
References
1. Ozhmegova E.N. et al. HIV drug resistance: past and current trends. Problems of Virology, 2022, vol. 67, no. 3, pp. 193–205. (In Russian) https://doi.org/10.36233/0507-4088-113
2. Xun J. et al. Editing out HIV: application of gene editing technology to achieve functional cure. Retrovirology, 2021, vol. 18, no. 1, pp. 39. https://doi.org/10.1186/s12977-021-00581-1
3. Del Corpo O. et al. A U1i RNA that enhances HIV-1 RNA splicing with an elongated recognition domain is an optimal candidate for combination HIV-1 gene therapy. Molecular Therapy-Nucleic Acids, 2019, vol. 18, pp. 815–830. https://doi.org/10.1016/j.omtn.2019.10.011
4. Virgilio A. et al. Novel monomolecular derivatives of the anti-HIV-1 G-quadruplex-forming Hotoda’s aptamer containing inversion of polarity sites. European Journal of Medicinal Chemistry, 2020, vol. 208, article id: 112786. https://doi.org/10.1016/j.ejmech.2020.112786
5. Cena-Diez R. et al. Novel naturally occurring dipeptides and single-stranded oligonucleotide act as entry inhibitors and exhibit a strong synergistic anti-HIV-1 profile. Infectious Diseases and Therapy, 2022, vol. 11, no. 3, pp. 1103–1116. https://doi.org/10.1007/s40121-022-00626-8
6. Torbett B., Goodsell D., Richman D. Targeting the HIV RNA Genome: High-Hanging Fruit Only Needs a Longer Ladder. Current Topics in Microbiology and Immunology, vol. 389, pp. 147–169. https://doi.org/10.1007/82_2015_434
7. Virgilio A. et al. Novel monomolecular derivatives of the anti-HIV-1 G-quadruplex-forming Hotoda’s aptamer containing inversion of polarity sites. European Journal of Medicinal Chemistry, 2020, vol. 208, article id: 112786.https://doi.org/10.1016/j.ejmech.2020.112786
8. Pashkov E.A., Pak A.V., Pashkov E.P., Bykov A.S., Budanova E.V., Poddubikov A.V., Svitich O.A., Zverev V.V. Perspektiva primeneniya preparatov na osnove yavleniya RNK-interferentsii protiv HIV-infection. Voprosy virusologii, 2022, vol. 67, no. 4, pp. 278–289. (In Russian) https://doi.org/10.36233/0507-4088-124
9. Lj R. A simple method of estimating fifty per cent endpoints. Am J Hyg, 1938, vol. 27, pp. 493–495. https://doi.org/10.1093/oxfordjournals.aje.a118408
10. Markov O.V. et al. Transport Oligonucleotides—A Novel System for Intracellular Delivery of Antisense Therapeutics. Molecules, 2020, vol. 25, no. 16, article id: 3663. https://doi.org/10.3390/molecules25163663
11. Kupryushkin M.S., Pyshnyi D.V., Stetsenko D.A. Phosphoryl guanidines: a new type of nucleic acid analogues. Acta Naturae, 2014, vol. 6, no. 4 (23), pp. 116–118. https://doi.org/10.32607/20758251-2014-6-4-116-118
12. Lomzov A.A. et al. Diastereomers of a monosubstituted phosphoryl guanidine trideoxyribonucleotide: isolation and properties. Biochemical and biophysical research communications, 2019, vol. 513, no. 4, pp. 807–811. https://doi.org/10.1016/j.bbrc.2019.04.024
13. Crooke S.T. et al. The interaction of phosphorothioatecontaining RNA targeted drugs with proteins is a critical determinant of the therapeutic effects of these agents. Journal of the American Chemical Society, 2020, vol. 142, no. 35, pp. 14754–14771. https://doi.org/10.1021/jacs.0c04928
14. Crooke S.T., Vickers T.A., Liang X. Phosphorothioate modified oligonucleotide–protein interactions. Nucleic acids research, 2020, vol. 48, no. 10, pp. 5235–5253. https://doi.org/10.1093/nar/gkaa299
15. Juliano R.L. The delivery of therapeutic oligonucleotides. Nucleic acids research, 2016, vol. 44, no. 14, pp. 6518–6548. https://doi.org/10.1093/nar/gkw236
16. Graham M.J. et al. In vivo distribution and metabolism of a phosphorothioate oligonucleotide within rat liver after intravenous administration. Journal of Pharmacology and Experimental Therapeutics, 1998, vol. 286, no. 1, pp. 447–458.
17. Crooke S.T. et al. Cellular uptake and trafficking of antisense oligonucleotides. Nature biotechnology, 2017, vol. 35, no. 3, pp. 230–237. https://doi.org/10.1038/nbt.3779
18. Iversen P.L. et al. Cellular uptake and subcellular distribution of phosphorothioate oligonucleotides into cultured cells. Antisense research and development, 1992, vol. 2, no. 3, pp. 211–222.
19. Lorenz P. et al. Nucleocytoplasmic shuttling: a novel in vivo property of antisense phosphorothioate oligodeoxynucleotides. Nucleic Acids Research, 2000, vol. 28, no. 2, pp. 582–592. https://doi.org/10.1093/nar/28.2.582
20. Turner J.J., Arzumanov A.A., Gait M.J. Synthesis, cellular uptake and HIV-1 Tat-dependent trans-activation inhibition activity of oligonucleotide analogues disulphide-conjugated to cell-penetrating peptides. Nucleic acids research, 2005, vol. 33, no. 1, pp. 27–42. https://doi.org/10.1093/nar/gki142
21. Takahashi M. et al. Dual mechanisms of action of selfdelivering, anti-HIV-1 FANA oligonucleotides as a potential new approach to HIV therapy. Molecular Therapy-Nucleic Acids, 2019, vol. 17, pp. 615–625. https://doi.org/10.1016/j.omtn.2019.07.001
22. Zhou J., Satheesan S., Li H., Weinberg M.S., Morris K.V., Burnett J.C., Rossi J.J. Cell-Specific RNA Aptamer against Human CCR5 Specifically Targets HIV-1 Susceptible Cells and Inhibits HIV-1 Infectivity. Chemistry & Biology, 2015, vol. 22, no. 3, pp 379-390. https://doi.org/10.1016/j.chembiol.2015.01.005
Review
For citations:
Gotfrid L.G., Pavlova A.S., Kupryushkin M.S., Pyshnaya I.A., Gashnikova N.M. Antiviral activity of modified oligonucleotides in human lymphoid cells infected with a strain of HIV‐1. South of Russia: ecology, development. 2024;19(4):57-67. (In Russ.) https://doi.org/10.18470/1992-1098-2024-4-5
Views:
116
Email this article 
