Response of murine neural stem/progenitor cells to gamma-neutron radiation

Abstract

Purpose

In recent years, a growing number of studies have focused on the mechanisms of action of densely ionizing radiation. This is associated with the development of radiation therapy of tumors using accelerated ions. The use of densely ionizing radiation appears to be the most promising method, optimal for treating patients with severe radioresistant forms, such as widespread head and neck tumors, recurrent and metastatic tumors, and some forms of brain tumors. The goal of our study was to investigate the effects of gamma-neutron radiation on mouse neural stem/progenitor cells (NSCs/NPCs).

Methods

NSCs/NPCs were isolated from neonatal mouse brains. Cells were irradiated in a collimated beam of neutrons and gamma rays of the IR-8 nuclear reactor. At 5 and 7 days after irradiation, cells and neurospheres were counted to assess survival. The number of DNA double-strand breaks and their repair efficiency were determined by immunocytochemical γH2AX staining followed by counting the number of γH2AX foci using a fluorescent microscope.

Results

We observed a dose-dependent decrease in the survival of NSCs/NPCs after irradiation at doses above 100 mGy and stimulation of the proliferation of these cells at doses of 25 and 50 mGy. In terms of a decrease in cell survival, the effect of gamma-neutron irradiation significantly exceeded the effect of gamma irradiation: the maximum value of the relative biological efficiency for gamma-neutron irradiation comprised 9.7. Gamma-neutron irradiation led to the formation of double-strand DNA breaks detected by the formation of foci of histone γH2AX in the cell nuclei. The γH2AX foci formed after gamma-neutron irradiation of NSCs/NPCs at doses of 100–500 mGy were characterized by a larger size in comparison with foci induced by gamma irradiation and gamma-neutron irradiation at a dose of 50 mGy. The repair of double-strand DNA breaks induced by γ,n-irradiation was slow; the repair rate depended on the radiation dose.

Conclusions

The data obtained indicate high sensitivity of proliferating NSCs/NPCs to gamma-neutron radiation. High RBE of gamma-neutron radiation requires special measures to protect the neurogenic regions of the brain when using this type of radiation in radiation therapy.

Keywords:

• Neural stem cells
• double-strand DNA breaks
• DNA repair
• γH2AX
• gamma-neutron radiation

Acknowledgments

The authors are grateful to Prof. E. Yu. Moskaleva for discussion and critical comments in the process of work on the manuscript, Yu. N. Panin for help in cells irradiation.

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

This work was supported by the National Research Center ‘Kurchatov Institute’ (order No. 1059 of 2 July 2020).

Notes on contributors

Galina A. Posypanova

Galina A. Posypanova, Ph.D., Dr. Sci, is a leading researcher at the Department of Cell Biology, Immunology and Molecular Medicine, National Research Center ‘Kurchatov Institute’ and professor at Faculty of Immunological Chemistry, Russian Technological University, Moscow, Russia.

Marya G. Ratushnyak

Marya G. Ratushnyak, Ph.D., is a researcher at the Department of Cell Biology, Immunology and Molecular Medicine, National Research Center ‘Kurchatov Institute’, Russia.

Yuliya P. Semochkina

Yuliya P. Semochkina, M.Sc., is a researcher at the Department of Cell Biology, Immunology and Molecular Medicine, National Research Center ‘Kurchatov Institute’, Russia.

Alexander N. Strepetov

Alexander N. Strepetov, Ph.D., is a senior researcher in Kurchatov Nuclear Physics Complex, National Research Center ‘Kurchatov Institute’, Russia.