Assessment of the preventive effect of knockdown of cellular genes NXF1, PRPS1PRPS1 and NAA10 in influenza infection in an in vitro model

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Abstract

Introduction. Influenza is an acute respiratory viral infectious disease caused by the influenza viruses. Current preventive and therapeutic approaches are of great anti-epidemic importance, but there are a number of problems, such as the rapid emergence of resistant strains, the lack of cross-immunity and the effectiveness of vaccines. One of the approaches to the development of anti-influenza agents is the use of RNA interference and small interfering RNAs complementary to the mRNA target of viral and cellular genes.

Aim ‒ to evaluate the prophylactic anti-influenza effect of siRNAs directed to the cellular genes NXF1, PRPS1 and NAA10 in an in vitro model.

Materials and methods. Antigenic variants of influenza A virus: A/California/7/09 (H1N1), A/WSN/33 (H1N1) and A/Brisbane/59/07 (H1N1); cell cultures A549 and MDCK. The study was performed using molecular genetic (transfection, NC isolation, RT-PCR-RV) and virological (cell culture infection, titration by visual CPE, viral titer assessment using the Ramakrishnan method) methods.

Results. It was shown that siRNAs targeting the cellular genes NXF1, PRPS1 and NAA10, when used prophylactically in cell culture at a concentration of 0.25 μg per well, during infection with influenza virus strains A/California/7/09 (H1N1), A/WSN/33 (H1N1) and A/Brisbane/59/07 (H1N1) at a multiplicity of infection of 0.01, reduced viral replication to a level of 220 TCID50 per 1 ml of cell medium, whereas in control untreated cells the viral yield was ~106 TCID50 per 1 ml of medium.

Conclusions. Reproduction of influenza A viruses directly depends on the protein products of the NXF1, PRPS1, and NAA10 genes. Reduced expression of these genes disrupts the life cycle and activity of influenza viruses. Such an approach can potentially be studied and used for closely and distantly related representatives of other virus families.

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

Ph. D., assistant of microbiology, virology and immunology department of Sechenov University; junior researcher laboratory of virology applied of I. Mechnikov Research Institute of Vaccines and Sera

Russian Federation, 119991, Moscow; 105064, Moscow

Dmitry A. Shikvin

Moscow State University of Fine Chemical Technologies

Email: carrypool@gmail.com
ORCID iD: 0009-0005-9874-2081

student of the Department of Biotechnology and Industrial Pharmacy of the Institute of Fine Chemical Technologies named after M.V. Lomonosov

Russian Federation, 119454, Moscow

George 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

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

student of the Institute of Children Health

Russian Federation, 119991, Moscow; 105064, Moscow

Firaya G. Nagieva

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

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

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

Russian Federation, 119991, Moscow

Ekaterina A. Bogdanova

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

Email: bogdekaterin@yandex.ru
ORCID iD: 0000-0002-5620-1843

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

Russian Federation, 105064, Moscow

Anatoly S. Bykov

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

Email: drbykov@bk.ru
ORCID iD: 0000-0002-8099-6201

MD, Professor of Microbiology, Virology and Immunology department

Russian Federation, 105064, Moscow

Evgeny P. Pashkov

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

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

MD, Professor of Microbiology, Virology and Immunology department

Russian Federation, 105064, 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 of I. Mechnikov Research Institute of Vaccines and Sera; Professor of Microbiology, Virology and Immunology department of Sechenov University

Russian Federation, 119991, Moscow; 105064, Moscow

Vitaly 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 of I. Mechnikov Research Institute of Vaccines and Sera; Professor, The Leader of Microbiology, Virology and Immunology department of Sechenov University

Russian Federation, 119991, Moscow; 105064, Moscow

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2. Fig. 1. Dynamic changes in NXF1, PRPS1 and NAA10 gene transcripts. siRNA and their target cellular genes of the same name are shown on the X-axis; % change in transcript level is shown on the Y-axis. Pfaffl test data are presented in %. * ‒ p ≤ 0.05.

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3. Fig. 2. Antiviral effect of siRNAs specific to the NXF1, PRPS1 and NAA10 cell genes. а ‒ A/California/7/09 (H1N1); b ‒ A/Brisbane/59/07 (H1N1); c ‒ A/WSN/1933 (H1N1); on the X-axis ‒ siRNA and their target cellular genes of the same name; on the Y-axis ‒ the viral titer log10 TCID50/mL relative to the viral and nonspecific control). * ‒ p < 0.05 relative to the nonspecific control siL2.

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Copyright (c) 2025 Pashkov E.A., Shikvin D.A., Pashkov G.A., Nagieva F.G., Bogdanova E.A., Bykov A.S., Pashkov E.P., Svitich O.A., Zverev V.V.

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