Characterization of biological peculiarities of the radioprotective activity of double-stranded RNA isolated from Saccharomyces сerevisiae

Abstract

The purpose of the article

Protection from ionizing radiation is the most important component in the curing malignant neoplasms, servicing atomic reactors, and resolving the situations associated with uncontrolled radioactive pollutions. In this regard, discovering new effective radioprotectors as well as novel principles of protecting living organisms from high-dose radiation is the most important factor, determining the new approaches in medical and technical usage of radiation.

Materials and methods

Experimental animals were irradiated on the γ-emitter (Cs¹³⁷) with a dose of 9.4 Gy. Radioprotective properties of several agents (total RNA, single-stranded RNA, double-stranded RNA and B-190) were estimated by the survival/death rates of experimental animals within 30–90 d. Pathomorphological examination of internal organs end electron microscope assay was done on days 9–12 after irradiation. Cloning and other molecular procedures were performed accordingly to commonly accepted protocols. For assessment of the internalization of labeled nucleic acid, bone marrow cells were incubated with double-stranded RNA labeled with 6-FAM fluorescent dye. Cells with internalized double-stranded RNA were assayed using Axio Imager M1 microscope. In the other experiment, bone marrow cells after incubation with double-stranded RNA were stained with Cy5-labeled anti-CD34 antibodies and assayed using Axioskop 2 microscope.

Results

In this study, several biological features of the radioprotective action of double-stranded RNA are characterized. It was shown that 160 µg of the double-stranded RNA per mouse protect experimental animals from the absolutely lethal dose of γ-radiation of 9.4 Gy. In different experiments, 80–100% of irradiated animals survive and live until their natural death. Radioprotective properties of double-stranded RNA were found to be independent on its sequence, but strictly dependent on its double-stranded form. Moreover, double-stranded RNA must have ‘open’ ends of the molecule to exert its radioprotective activity.

Conclusions

Experiments indicate that radioprotective effect of double-stranded RNA is tightly bound to its internalization into hematopoietic stem cells, which further repopulate the spleen parenchyma of irradiated mice. Actively proliferating progenitors form the splenic colonies, which further serve as the basis for restoration of hematopoiesis and immune function and determine the survival of animals received the lethal dose of radiation.

Keywords:

• Double-stranded RNA
• B-190
• spleen colonies
• double-strand breaks

Acknowledgments

The authors express their gratitude to Prof. Sergey V. Netesov for technical support, the laboratory of immunogenetics of the Institute of Molecular and Cellular Biology SB RAS for providing XL1-Blue MRF cells, the Radioisotope Division of ICG SB RAS, the Flow Cytometry Shared Facility ICG SB RAS, and the Common Use Center for Microscopy of Biologic Objects SB RAS.

Disclosure statement

The authors report no conflict of interest.

Additional information

Funding

The work was supported by the Russian Ministry of Science and High Education via the Institute of Cytology and Genetics [state budget project No 0324-2019-0042-C-01, state registration No AAAA-A17-117071240065-4] and Russian Foundation for Basic Research [grant No 18-34-00205]. Microscopy analysis was supported by the Russian State-funded budget project of the Institute of Molecular and Cellular Biology SB RAS [No 0310-2019-0005]. The RNA’s chemical synthesis work was supported by the Russian State-funded budget project of the Institute of Chemical Biology and Fundamental Medicine SB RAS [No AAAA-A17-117020210021-7].

Notes on contributors

Genrikh S. Ritter

Genrikh S. Ritter have graduated from Novosibirsk State University. Now he is a PhD student and Researcher of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.

Valeriy P. Nikolin

Valeriy P. Nikolin have graduated from Novosibirsk State Medical University. He is a PhD and has many years of experience working with experimental animals.

Nelly A. Popova

Nelly A. Popova have graduated from Novosibirsk State Medical University. She is a PhD and Professor of Novosibirsk State University.

Anastasia S. Proskurina

Anastasia S. Proskurina have graduated from Novosibirsk State University. She is a PhD and the Senior Research Associate of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.

Polina E. Kisaretova

Polina E. Kisaretova have graduated from Novosibirsk State University. Now she is a PhD student.

Oleg S. Taranov

Oleg S. Taranov is the Head of the Department of Microscopic Research in the State Research Center of Virology and Biotechnology “Vector”. He has many years of experience in pathomorphological assay and electron microscopy.

Tatiana D. Dubatolova

Tatiana D. Dubatolova is a Researcher of Institute of Molecular and Cellular Biology and has many years of experience in microscopic analysis of blood cells.

Evgenia V. Dolgova

Evgenia V. Dolgova have graduated from Novosibirsk State University. She is a PhD and the Senior Research Associate of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.

Ekaterina A. Potter

Ekaterina A. Potter have graduated from Novosibirsk State University. She is a PhD and the Senior Research Associate of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.

Svetlana S. Kirikovich

Svetlana S. Kirikovich is a PhD and the Research Associate of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.

Yaroslav R. Efremov

Yaroslav R. Efremov have graduated from Novosibirsk State University. He is a PhD and an Engineer at the Flow Cytometry Shared Facility ICG SB RAS.

Sergey I. Bayborodin

Sergey I. Bayborodin have graduated from Novosibirsk State University. He is a PhD and the head of the Common Use Center for Microscopy of Biologic Objects SB RAS.

Margarita V. Romanenko

Margarita V. Romanenko have graduated from Novosibirsk State University. She is a PhD and the Research Associate of the Novosibirsk State University.

Maria I. Meschaninova

Maria I. Meschaninova is a PhD and the Research Associate of the RNA Chemistry Laboratory in the Institute of Chemical Biology and Fundamental Medicine.

Aliya G. Venyaminova

Aliya G. Venyaminova is a PhD and the Head of the RNA Chemistry Laboratory in the Institute of Chemical Biology and Fundamental Medicine.

Nikolay A. Kolchanov

Nikolay A. Kolchanov have graduated from Novosibirsk State University. He was the Director of the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences from 2008 to 2018 years. He is an Academician of the Russian Academy of Sciences and professor of Novosibirsk State University.

Mikhail A. Shurdov

Mikhail A. Shurdov have graduated from Novosibirsk State University. He is a PhD and the head of the Joint Stock Company “Cheboksary Electrical Apparatus Plant”.

Sergey S. Bogachev

Sergey S. Bogachev have graduated from Saratov State University. He is a Doctor of Science and the head of the Laboratory of induced cellular processes in the Institute of Cytology and Genetics.