DNA Damage and Repair in Peripheral Blood Mononuclear Cells After Internal Ex Vivo Irradiation of Patient Blood with I

Overview

Journal Eur J Nucl Med Mol Imaging

Specialties Biophysics
Nuclear Medicine
Radiology

Date 2021 Nov 13

PMID 34773472

Citations 12

Authors

S Schumann

H Scherthan

K Pfestroff

S Schoof

A Pfestroff

P Hartrampf

N Hasenauer

A K Buck

M Luster

M Port

M Lassmann

U Eberlein

Affiliations

Soon will be listed here.

Abstract

Aim: The aim of this study was to provide a systematic approach to characterize DNA damage induction and repair in isolated peripheral blood mononuclear cells (PBMCs) after internal ex vivo irradiation with [I]NaI. In this approach, we tried to mimic ex vivo the irradiation of patient blood in the first hours after radioiodine therapy.

Material And Methods: Blood of 33 patients of two centres was collected immediately before radioiodine therapy of differentiated thyroid cancer (DTC) and split into two samples. One sample served as non-irradiated control. The second sample was exposed to ionizing radiation by adding 1 ml of [I]NaI solution to 7 ml of blood, followed by incubation at 37 °C for 1 h. PBMCs of both samples were isolated, split in three parts each and (i) fixed in 70% ethanol and stored at - 20 °C directly (0 h) after irradiation, (ii) after 4 h and (iii) 24 h after irradiation and culture in RPMI medium. After immunofluorescence staining microscopically visible co-localizing γ-H2AX + 53BP1 foci were scored in 100 cells per sample as biomarkers for radiation-induced double-strand breaks (DSBs).

Results: Thirty-two of 33 blood samples could be analysed. The mean absorbed dose to the blood in all irradiated samples was 50.1 ± 2.3 mGy. For all time points (0 h, 4 h, 24 h), the average number of γ-H2AX + 53BP1 foci per cell was significantly different when compared to baseline and the other time points. The average number of radiation-induced foci (RIF) per cell after irradiation was 0.72 ± 0.16 at t = 0 h, 0.26 ± 0.09 at t = 4 h and 0.04 ± 0.09 at t = 24 h. A monoexponential fit of the mean values of the three time points provided a decay rate of 0.25 ± 0.05 h, which is in good agreement with data obtained from external irradiation with γ- or X-rays.

Conclusion: This study provides novel data about the ex vivo DSB repair in internally irradiated PBMCs of patients before radionuclide therapy. Our findings show, in a large patient sample, that efficient repair occurs after internal irradiation with 50 mGy absorbed dose, and that the induction and repair rate after I exposure is comparable to that of external irradiation with γ- or X-rays.

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References

Aerts A, Eberlein U, Holm S, Hustinx R, Konijnenberg M, Strigari L . EANM position paper on the role of radiobiology in nuclear medicine. Eur J Nucl Med Mol Imaging. 2021; 48(11):3365-3377. PMC: 8440244. DOI: 10.1007/s00259-021-05345-9. View

Eberlein U, Nowak C, Bluemel C, Buck A, Werner R, Scherthan H . DNA damage in blood lymphocytes in patients after (177)Lu peptide receptor radionuclide therapy. Eur J Nucl Med Mol Imaging. 2015; 42(11):1739-1749. PMC: 4554740. DOI: 10.1007/s00259-015-3083-9. View

Eberlein U, Scherthan H, Bluemel C, Peper M, Lapa C, Buck A . DNA Damage in Peripheral Blood Lymphocytes of Thyroid Cancer Patients After Radioiodine Therapy. J Nucl Med. 2015; 57(2):173-9. DOI: 10.2967/jnumed.115.164814. View

Schumann S, Eberlein U, Muhtadi R, Lassmann M, Scherthan H . DNA damage in leukocytes after internal ex-vivo irradiation of blood with the α-emitter Ra-223. Sci Rep. 2018; 8(1):2286. PMC: 5797089. DOI: 10.1038/s41598-018-20364-7. View

Ivashkevich A, Redon C, Nakamura A, Martin R, Martin O . Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research. Cancer Lett. 2011; 327(1-2):123-33. PMC: 3329565. DOI: 10.1016/j.canlet.2011.12.025. View

Lamkowski A, Forcheron F, Agay D, Ahmed E, Drouet M, Meineke V . DNA damage focus analysis in blood samples of minipigs reveals acute partial body irradiation. PLoS One. 2014; 9(2):e87458. PMC: 3911974. DOI: 10.1371/journal.pone.0087458. View

Panier S, Boulton S . Double-strand break repair: 53BP1 comes into focus. Nat Rev Mol Cell Biol. 2013; 15(1):7-18. DOI: 10.1038/nrm3719. View

Zimmermann M, de Lange T . 53BP1: pro choice in DNA repair. Trends Cell Biol. 2013; 24(2):108-17. PMC: 3946699. DOI: 10.1016/j.tcb.2013.09.003. View

Lassmann M, Hanscheid H, Gassen D, Biko J, Meineke V, Reiners C . In vivo formation of gamma-H2AX and 53BP1 DNA repair foci in blood cells after radioiodine therapy of differentiated thyroid cancer. J Nucl Med. 2010; 51(8):1318-25. DOI: 10.2967/jnumed.109.071357. View

10.

Eberlein U, Peper M, Fernandez M, Lassmann M, Scherthan H . Calibration of the γ-H2AX DNA double strand break focus assay for internal radiation exposure of blood lymphocytes. PLoS One. 2015; 10(4):e0123174. PMC: 4390303. DOI: 10.1371/journal.pone.0123174. View

11.

Horn S, Brady D, Prise K . Alpha particles induce pan-nuclear phosphorylation of H2AX in primary human lymphocytes mediated through ATM. Biochim Biophys Acta. 2015; 1853(10 Pt A):2199-206. DOI: 10.1016/j.bbamcr.2015.06.010. View

12.

Chowdhury D, Keogh M, Ishii H, Peterson C, Buratowski S, Lieberman J . gamma-H2AX dephosphorylation by protein phosphatase 2A facilitates DNA double-strand break repair. Mol Cell. 2005; 20(5):801-9. DOI: 10.1016/j.molcel.2005.10.003. View

13.

Schumann S, Eberlein U, Muller J, Scherthan H, Lassmann M . Correlation of the absorbed dose to the blood and DNA damage in leukocytes after internal ex-vivo irradiation of blood samples with Ra-224. EJNMMI Res. 2018; 8(1):77. PMC: 6082247. DOI: 10.1186/s13550-018-0422-4. View

14.

Lobrich M, Rief N, Kuhne M, Heckmann M, Fleckenstein J, Rube C . In vivo formation and repair of DNA double-strand breaks after computed tomography examinations. Proc Natl Acad Sci U S A. 2005; 102(25):8984-9. PMC: 1150277. DOI: 10.1073/pnas.0501895102. View

15.

Beels L, Werbrouck J, Thierens H . Dose response and repair kinetics of gamma-H2AX foci induced by in vitro irradiation of whole blood and T-lymphocytes with X- and gamma-radiation. Int J Radiat Biol. 2010; 86(9):760-8. DOI: 10.3109/09553002.2010.484479. View

16.

Grudzenski S, Raths A, Conrad S, Rube C, Lobrich M . Inducible response required for repair of low-dose radiation damage in human fibroblasts. Proc Natl Acad Sci U S A. 2010; 107(32):14205-10. PMC: 2922519. DOI: 10.1073/pnas.1002213107. View

17.

Horn S, Barnard S, Rothkamm K . Gamma-H2AX-based dose estimation for whole and partial body radiation exposure. PLoS One. 2011; 6(9):e25113. PMC: 3179476. DOI: 10.1371/journal.pone.0025113. View

18.

Mariotti L, Pirovano G, Savage K, Ghita M, Ottolenghi A, Prise K . Use of the γ-H2AX assay to investigate DNA repair dynamics following multiple radiation exposures. PLoS One. 2013; 8(11):e79541. PMC: 3843657. DOI: 10.1371/journal.pone.0079541. View

19.

van Oorschot B, Hovingh S, Dekker A, Stalpers L, Franken N . Predicting Radiosensitivity with Gamma-H2AX Foci Assay after Single High-Dose-Rate and Pulsed Dose-Rate Ionizing Irradiation. Radiat Res. 2016; 185(2):190-8. DOI: 10.1667/RR14098.1. View

20.

Lengert N, Mirsch J, Weimer R, Schumann E, Haub P, Drossel B . AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair. Sci Rep. 2018; 8(1):17282. PMC: 6251879. DOI: 10.1038/s41598-018-35660-5. View