Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact
Aptamers are currently being investigated for their potential to improve virotherapy. They offer several advantages, including the ability to prevent the aggregation of viral particles, enhance target specificity, and protect against the neutralizing effects of antibodies. The purpose of this study...
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MDPI AG
2024-02-01
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| author | Maya A. Dymova Daria O. Malysheva Victoria K. Popova Elena V. Dmitrienko Anton V. Endutkin Danil V. Drokov Vladimir S. Mukhanov Arina A. Byvakina Galina V. Kochneva Polina V. Artyushenko Irina A. Shchugoreva Anastasia V. Rogova Felix N. Tomilin Anna S. Kichkailo Vladimir A. Richter Elena V. Kuligina |
| author_facet | Maya A. Dymova Daria O. Malysheva Victoria K. Popova Elena V. Dmitrienko Anton V. Endutkin Danil V. Drokov Vladimir S. Mukhanov Arina A. Byvakina Galina V. Kochneva Polina V. Artyushenko Irina A. Shchugoreva Anastasia V. Rogova Felix N. Tomilin Anna S. Kichkailo Vladimir A. Richter Elena V. Kuligina |
| author_sort | Maya A. Dymova |
| collection | DOAJ |
| description | Aptamers are currently being investigated for their potential to improve virotherapy. They offer several advantages, including the ability to prevent the aggregation of viral particles, enhance target specificity, and protect against the neutralizing effects of antibodies. The purpose of this study was to comprehensively investigate an aptamer capable of enhancing virotherapy. This involved characterizing the previously selected aptamer for vaccinia virus (VACV), evaluating the aggregation and molecular interaction of the optimized aptamers with the recombinant oncolytic virus VV-GMCSF-Lact, and estimating their immunoshielding properties in the presence of human blood serum. We chose one optimized aptamer, NV14t_56, with the highest affinity to the virus from the pool of several truncated aptamers and built its 3D model. The NV14t_56 remained stable in human blood serum for 1 h and bound to VV-GMCSF-Lact in the micromolar range (Kd ≈ 0.35 μM). Based on dynamic light scattering data, it has been demonstrated that aptamers surround viral particles and inhibit aggregate formation. In the presence of serum, the hydrodynamic diameter (by intensity) of the aptamer–virus complex did not change. Microscale thermophoresis (MST) experiments showed that NV14t_56 binds with virus (EC50 = 1.487 × 10<sup>9</sup> PFU/mL). The analysis of the amplitudes of MST curves reveals that the components of the serum bind to the aptamer–virus complex without disrupting it. In vitro experiments demonstrated the efficacy of VV-GMCSF-Lact in conjunction with the aptamer when exposed to human blood serum in the absence of neutralizing antibodies (Nabs). Thus, NV14t_56 has the ability to inhibit virus aggregation, allowing VV-GMCSF-Lact to maintain its effectiveness throughout the storage period and subsequent use. When employing aptamers as protective agents for oncolytic viruses, the presence of neutralizing antibodies should be taken into account. |
| first_indexed | 2024-03-07T22:19:50Z |
| format | Article |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-07T22:19:50Z |
| publishDate | 2024-02-01 |
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| spelling | doaj.art-69ca6d94648b4369a5bacbc2e08347ad2024-02-23T15:29:04ZengMDPI AGMolecules1420-30492024-02-0129484810.3390/molecules29040848Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-LactMaya A. Dymova0Daria O. Malysheva1Victoria K. Popova2Elena V. Dmitrienko3Anton V. Endutkin4Danil V. Drokov5Vladimir S. Mukhanov6Arina A. Byvakina7Galina V. Kochneva8Polina V. Artyushenko9Irina A. Shchugoreva10Anastasia V. Rogova11Felix N. Tomilin12Anna S. Kichkailo13Vladimir A. Richter14Elena V. Kuligina15Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaState Research Center of Virology and Biotechnology “Vector”, 630559 Koltsovo, RussiaLaboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University Named after Prof. V.F. Voyno-Yasenetsky, Partizana Zheleznyaka str. 1, 660022 Krasnoyarsk, RussiaLaboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University Named after Prof. V.F. Voyno-Yasenetsky, Partizana Zheleznyaka str. 1, 660022 Krasnoyarsk, RussiaLaboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University Named after Prof. V.F. Voyno-Yasenetsky, Partizana Zheleznyaka str. 1, 660022 Krasnoyarsk, RussiaFederal Research Center KSC SB RAS, 50 Akademgorodok, 660036 Krasnoyarsk, RussiaLaboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University Named after Prof. V.F. Voyno-Yasenetsky, Partizana Zheleznyaka str. 1, 660022 Krasnoyarsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, RussiaAptamers are currently being investigated for their potential to improve virotherapy. They offer several advantages, including the ability to prevent the aggregation of viral particles, enhance target specificity, and protect against the neutralizing effects of antibodies. The purpose of this study was to comprehensively investigate an aptamer capable of enhancing virotherapy. This involved characterizing the previously selected aptamer for vaccinia virus (VACV), evaluating the aggregation and molecular interaction of the optimized aptamers with the recombinant oncolytic virus VV-GMCSF-Lact, and estimating their immunoshielding properties in the presence of human blood serum. We chose one optimized aptamer, NV14t_56, with the highest affinity to the virus from the pool of several truncated aptamers and built its 3D model. The NV14t_56 remained stable in human blood serum for 1 h and bound to VV-GMCSF-Lact in the micromolar range (Kd ≈ 0.35 μM). Based on dynamic light scattering data, it has been demonstrated that aptamers surround viral particles and inhibit aggregate formation. In the presence of serum, the hydrodynamic diameter (by intensity) of the aptamer–virus complex did not change. Microscale thermophoresis (MST) experiments showed that NV14t_56 binds with virus (EC50 = 1.487 × 10<sup>9</sup> PFU/mL). The analysis of the amplitudes of MST curves reveals that the components of the serum bind to the aptamer–virus complex without disrupting it. In vitro experiments demonstrated the efficacy of VV-GMCSF-Lact in conjunction with the aptamer when exposed to human blood serum in the absence of neutralizing antibodies (Nabs). Thus, NV14t_56 has the ability to inhibit virus aggregation, allowing VV-GMCSF-Lact to maintain its effectiveness throughout the storage period and subsequent use. When employing aptamers as protective agents for oncolytic viruses, the presence of neutralizing antibodies should be taken into account.https://www.mdpi.com/1420-3049/29/4/848aptameroncolytic virusgliomadynamic light scatteringmicroscale thermophoresis |
| spellingShingle | Maya A. Dymova Daria O. Malysheva Victoria K. Popova Elena V. Dmitrienko Anton V. Endutkin Danil V. Drokov Vladimir S. Mukhanov Arina A. Byvakina Galina V. Kochneva Polina V. Artyushenko Irina A. Shchugoreva Anastasia V. Rogova Felix N. Tomilin Anna S. Kichkailo Vladimir A. Richter Elena V. Kuligina Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact Molecules aptamer oncolytic virus glioma dynamic light scattering microscale thermophoresis |
| title | Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact |
| title_full | Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact |
| title_fullStr | Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact |
| title_full_unstemmed | Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact |
| title_short | Characterizing Aptamer Interaction with the Oncolytic Virus VV-GMCSF-Lact |
| title_sort | characterizing aptamer interaction with the oncolytic virus vv gmcsf lact |
| topic | aptamer oncolytic virus glioma dynamic light scattering microscale thermophoresis |
| url | https://www.mdpi.com/1420-3049/29/4/848 |
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