Influence of siRNA complexes on the reproduction of influenza A virus (Orthomyxoviridae: Alphainfluenzavirus) in vivo

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Abstract

Introduction. Influenza is one of the most pressing global health problems. Despite the wide range of available anti-influenza drugs, the viral drug resistance is an increasing concern and requires the search for new approaches to overcome it. A promising solution is the development of drugs with action that is based on the inhibition of the activity of cellular genes through RNA interference.

Aim. Evaluation in vivo of the preventive potential of miRNAs directed to the cellular genes FLT4, Nup98 and Nup205 against influenza infection.

Materials and methods. The A/California/7/09 strain of influenza virus (H1N1) and BALB/c mice were used in the study. The administration of siRNA and experimental infection of animals were performed intranasally. The results of the experiment were analyzed using molecular genetic and virological methods.

Results. The use of siRNA complexes Nup98.1 and Nup205.1 led to a significant decrease in viral reproduction and concentration of viral RNA on the 3rd day after infection. When two siRNA complexes (Nup98.1 and Nup205.1) were administered simultaneously, a significant decrease in viral titer and concentration of viral RNA was also noted compared with the control groups.

Conclusions. The use of siRNAs in vivo can lead to an antiviral effect when the activity of single or several cellular genes is suppressed. The results indicate that the use of siRNAs targeting the cellular genes whose expression products are involved in viral reproduction is one of the promising methods for the prevention and treatment of not only influenza, but also other respiratory infections.

About the authors

Evgeny A. Pashkov

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University); I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Author for correspondence.
Email: pashckov.j@yandex.ru
ORCID iD: 0000-0002-5682-4581

postgraduate of microbiology, virology and immunology department; junior researcher laboratory of virology applied

Russian Federation, 119991, Moscow; 105064, Moscow

Viktoriia Y. Momot

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: momot_v_yu@student.sechenov.ru
ORCID iD: 0000-0003-3476-5485

student of the Institute of Medical Biochemistry

Russian Federation, 119991, Moscow

Anastasia V. Pak

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: dcnnpk@gmail.com
ORCID iD: 0000-0003-4295-7858

student of the Institute of Clinical Medicine

Russian Federation, 119991, Moscow

Roman V. Samoilikov

I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: roma_sam78@mail.ru
ORCID iD: 0000-0001-6405-1390

researcher laboratory of molecular immunology

Russian Federation, 105064, Moscow

George A. Pashkov

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: georgp2004@mail.ru
ORCID iD: 0000-0003-0392-9969

student of the Institute of Children Health

Russian Federation, 119991, Moscow

Galina N. Usatova

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: g.n.usatova@mail.ru
ORCID iD: 0000-0002-8955-3570

Ph. D., private-docent Professor of Microbiology, Virology and Immunology department

Russian Federation, 119991, Moscow

Elena O. Kravtsova

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: elenakravtsov@yandex.ru
ORCID iD: 0000-0002-9100-0422

Ph. D., private-docent Professor of Microbiology, Virology and Immunology department

Russian Federation, 119991, Moscow

Alexander V. Poddubikov

I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: poddubikov@yandex.ru
ORCID iD: 0000-0001-8962-4765

Ph. D., The Head of laboratory of opportunistic pathogenic bacteria

Russian Federation, 105064, Moscow

Firaya G. Nagieva

I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: fgn42@yandex.ru
ORCID iD: 0000-0001-8204-4899

MD, private-docent, The Head of laboratory of hybrid cell cultures

Russian Federation, 105064, Moscow

Alexander V. Sidorov

I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: sashasidorov@yandex.ru
ORCID iD: 0000-0002-3561-8295

Ph. D., The Head of laboratory of DNA viruses

Russian Federation, 105064, Moscow

Evgeny P. Pashkov

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)

Email: 9153183256@mail.ru
ORCID iD: 0000-0002-4963-5053

MD, Professor of Microbiology, Virology and Immunology department

Russian Federation, 119991, Moscow

Oxana A. Svitich

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University); I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: svitichoa@yandex.ru
ORCID iD: 0000-0003-1757-8389

Corresponding member of RAS, MD, The head; Professor of Microbiology, Virology and Immunology department

Russian Federation, 119991, Moscow; 105064, Moscow

Vitaliy V. Zverev

Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University); I.I. Mechnikov Scientific and Research Institute of Vaccines and Sera

Email: vitalyzverev@outlook.com
ORCID iD: 0000-0002-0017-1892

Academician of RAS, Doctor of Biological Sciences, Scientific Adviser; Professor, The Leader of Microbiology, Virology and Immunology department

Russian Federation, 119991, Moscow; 105064, Moscow

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2. Fig. 1. Effect of siRNA on the expression of genes FLT4, Nup98, and Nup205.The X-axis shows the names of siRNAs targeting the genes of the same name. The Y-axis shows the changes in the expression of the FLT4, Nup98 and Nup205 genes after treatment of cells with siRNA complexes relative to nonspecific siRNA L2, expressed (in %). *р ≤ 0.05.

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3. Fig. 2. The effect of siRNA complexes on dynamic changes of mice body weight during three days after infection with the influenza A/California/7/09 (H1N1) virus.The X-axis shows siRNAs and respective target cellular genes of the same name. The Y-axis shows the changes in body weight, expressed (in %). *р ≤ 0.05.

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4. Fig. 3. Effect of siRNA suppression of the Nup98 and Nup205 genes on the reproduction of influenza A/California/7/09 (H1N1) virus on the third day after infection with LD50.The X-axis shows the name of the siRNA. The Y-axis shows the viral titers. *р ≤ 0.05

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5. Fig. 4. Effect of siRNA suppression of Nup98 and Nup205 genes on concentration of viral RNA on the third day after infection with LD50.The X-axis shows the names of siRNAs. The Y-axis shows the concentration of viral RNA. *р ≤ 0.05.

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6. Fig. 5. Effect of the Nup98.1/Nup205.1 siRNA pool on the reproduction of influenza A/California/7/09 (H1N1) virus on the third day after infection with LD50.The X-axis shows the names of the siRNAs. The Y-axis shows the viral titers. *р ≤ 0.05.

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7. Fig. 6. Effect of siRNA suppression of Nup98 and Nup205 genes on concentration of viral RNA on the third day after challenge with LD50.The X-axis shows the names of siRNAs. The Y-axis shows the concentration of viral RNA. *p < 0.05.

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Copyright (c) 2023 Pashkov E.A., Momot V.Y., Pak A.V., Samoilikov R.V., Pashkov G.A., Usatova G.N., Kravtsova E.O., Poddubikov A.V., Nagieva F.G., Sidorov A.V., Pashkov E.P., Svitich O.A., Zverev V.V.

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