Inter-laboratory comparison of gene expression biodosimetry for protracted radiation exposures as part of the RENEB and EURADOS WG10 2019 exercise.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
07 05 2021
07 05 2021
Historique:
received:
05
11
2020
accepted:
25
02
2021
entrez:
8
5
2021
pubmed:
9
5
2021
medline:
21
5
2022
Statut:
epublish
Résumé
Large-scale radiation emergency scenarios involving protracted low dose rate radiation exposure (e.g. a hidden radioactive source in a train) necessitate the development of high throughput methods for providing rapid individual dose estimates. During the RENEB (Running the European Network of Biodosimetry) 2019 exercise, four EDTA-blood samples were exposed to an Iridium-192 source (1.36 TBq, Tech-Ops 880 Sentinal) at varying distances and geometries. This resulted in protracted doses ranging between 0.2 and 2.4 Gy using dose rates of 1.5-40 mGy/min and exposure times of 1 or 2.5 h. Blood samples were exposed in thermo bottles that maintained temperatures between 39 and 27.7 °C. After exposure, EDTA-blood samples were transferred into PAXGene tubes to preserve RNA. RNA was isolated in one laboratory and aliquots of four blinded RNA were sent to another five teams for dose estimation based on gene expression changes. Using an X-ray machine, samples for two calibration curves (first: constant dose rate of 8.3 mGy/min and 0.5-8 h varying exposure times; second: varying dose rates of 0.5-8.3 mGy/min and 4 h exposure time) were generated for distribution. Assays were run in each laboratory according to locally established protocols using either a microarray platform (one team) or quantitative real-time PCR (qRT-PCR, five teams). The qRT-PCR measurements were highly reproducible with coefficient of variation below 15% in ≥ 75% of measurements resulting in reported dose estimates ranging between 0 and 0.5 Gy in all samples and in all laboratories. Up to twofold reductions in RNA copy numbers per degree Celsius relative to 37 °C were observed. However, when irradiating independent samples equivalent to the blinded samples but increasing the combined exposure and incubation time to 4 h at 37 °C, expected gene expression changes corresponding to the absorbed doses were observed. Clearly, time and an optimal temperature of 37 °C must be allowed for the biological response to manifest as gene expression changes prior to running the gene expression assay. In conclusion, dose reconstructions based on gene expression measurements are highly reproducible across different techniques, protocols and laboratories. Even a radiation dose of 0.25 Gy protracted over 4 h (1 mGy/min) can be identified. These results demonstrate the importance of the incubation conditions and time span between radiation exposure and measurements of gene expression changes when using this method in a field exercise or real emergency situation.
Identifiants
pubmed: 33963206
doi: 10.1038/s41598-021-88403-4
pii: 10.1038/s41598-021-88403-4
pmc: PMC8105310
doi:
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
9756Subventions
Organisme : NIAID NIH HHS
ID : U19 AI067773
Pays : United States
Références
Chaudhry, M. A. Biomarkers for human radiation exposure. J. Biomed. Sci. 15, 557–563 (2008).
doi: 10.1007/s11373-008-9253-z
Agency, I. A. E. Cytogenetic dosimetry: Applications in preparedness for and response to radiation emergencies. Man. Ser. 247 (2011).
Rothkamm, K. et al. Comparison of established and emerging biodosimetry assays. Radiat. Res. 180, 111–119 (2013).
doi: 10.1667/RR3231.1
Amundson, S. A. et al. Identification of potential mRNA biomarkers in peripheral blood lymphocytes for human exposure to ionizing radiation. Radiat. Res. 154, 342–346 (2000).
doi: 10.1667/0033-7587(2000)154[0342:IOPMBI]2.0.CO;2
Dressman, H. K. et al. Gene expression signatures that predict radiation exposure in mice and humans. PLoS Med. 4, 690–701 (2007).
doi: 10.1371/journal.pmed.0040106
Kultova, G., Tichy, A., Rehulkova, H. & Myslivcova-Fucikova, A. The hunt for radiation biomarkers: Current situation. Int. J. Radiat. Biol. 96, 370–382 (2020).
doi: 10.1080/09553002.2020.1704909
Badie, C. et al. Laboratory intercomparison of gene expression assays. Radiat. Res. 180, 138–148 (2013).
doi: 10.1667/RR3236.1
Port, M. et al. Rapid high-throughput diagnostic triage after a mass radiation exposure event using early gene expression changes. Radiat. Res. 192, 208–218 (2019).
doi: 10.1667/RR15360.1
Paul, S. & Amundson, S. A. Development of gene expression signatures for practical radiation biodosimetry. Int. J. Radiat. Oncol. Biol. Phys. 71, 1236–1244 (2008).
doi: 10.1016/j.ijrobp.2008.03.043
Brengues, M. et al. Biodosimetry on small blood volume using gene expression assay. Health Phys. 98, 179–185 (2010).
doi: 10.1097/01.HP.0000346706.44253.5c
Kabacik, S. et al. Gene expression following ionising radiation: Identification of biomarkers for dose estimation and prediction of individual response. Int. J. Radiat. Biol. 87, 115–129 (2011).
doi: 10.3109/09553002.2010.519424
Boldt, S., Knops, K., Kriehuber, R. & Wolkenhauer, O. A frequency-based gene selection method to identify robust biomarkers for radiation dose prediction. Int. J. Radiat. Biol. 88, 267–276 (2012).
doi: 10.3109/09553002.2012.638358
Knops, K., Boldt, S., Wolkenhauer, O. & Kriehuber, R. Gene expression in low- and high-dose-irradiated human peripheral blood lymphocytes: Possible applications for biodosimetry. Radiat. Res. 178, 304–312 (2012).
doi: 10.1667/RR2913.1
Cruz-Garcia, L. et al. Generation of a transcriptional radiation exposure signature in human blood using long-read nanopore sequencing. Radiat. Res. https://doi.org/10.1667/rr15476.1 (2019).
doi: 10.1667/rr15476.1
Cruz-Garcia, L. et al. In vivo validation of alternative fdxr transcripts in human blood in response to ionizing radiation. Int. J. Mol. Sci. 21, 1–18 (2020).
doi: 10.3390/ijms21217851
Bedford, J. S. & Mitchell, J. B. Dose rate effects in synchronous mammalian cells in culture. Radiat. Res. 54, 316–327 (1973).
doi: 10.2307/3573709
Hall, E. J. & Giaccia, A. J. Radiobiology for the radiologist: Seventh edition. Radiobiology for the Radiologist: Seventh Edition (2012).
Gridley, D. S., Rizvi, A., Luo-Owen, X., Makinde, A. Y. & Pecaut, M. J. Low dose, low dose rate photon radiation Modifies leukocyte distribution and gene expression in CD4+ T cells. J. Radiat. Res. 50, 139–150 (2009).
doi: 10.1269/jrr.08095
Paul, S. et al. Gene expression response of mice after a single dose of 137Cs as an internal emitter. Radiat. Res. 182, 380–389 (2014).
doi: 10.1667/RR13466.1
Ghandhi, S. A. et al. Effect of 90Sr internal emitter on gene expression in mouse blood. BMC Genomics 16, 1–15 (2015).
doi: 10.1186/s12864-015-1774-z
Ghandhi, S. A., Smilenov, L. B., Elliston, C. D., Chowdhury, M. & Amundson, S. A. Radiation dose-rate effects on gene expression for human biodosimetry. Functional and structural genomics. BMC Med. Genomics 8, 1–10 (2015).
doi: 10.1186/s12920-015-0097-x
Ghandhi, S. A., Shuryak, I., Morton, S. R., Amundson, S. A. & Brenner, D. J. New approaches for quantitative reconstruction of radiation dose in human blood cells. Sci. Rep. 9, 1–11 (2019).
doi: 10.1038/s41598-019-54967-5
Fachin, A. L. et al. Gene expression profiles in radiation workers occupationally exposed to ionizing radiation. J. Radiat. Res. 50, 61–71 (2009).
doi: 10.1269/jrr.08034
Rudqvist, N. et al. Transcriptional response in mouse thyroid tissue after 211at administration: Effects of absorbed dose, initial dose-rate and time after administration. PLoS ONE 10, e0131686 (2015).
doi: 10.1371/journal.pone.0131686
Morandi, E. et al. Gene expression changes in medical workers exposed to radiation. Radiat. Res. https://doi.org/10.1667/rr1545.1 (2009).
doi: 10.1667/rr1545.1
Abend, M. et al. Gene expression analysis in mayak workers with prolonged occupational radiation exposure. Health Phys. 106, 664–676 (2014).
doi: 10.1097/HP.0000000000000018
Shuryak, I. Review of resistance to chronic ionizing radiation exposure under environmental conditions in multicellular organisms. J. Environ. Radioact. 212, 106128 (2020).
doi: 10.1016/j.jenvrad.2019.106128
Wang, Q. et al. DNA damage response in peripheral mouse blood leukocytes in vivo after variable, low-dose rate exposure. Radiat. Environ. Biophys. 59, 89–98 (2020).
doi: 10.1007/s00411-019-00825-x
Gillies, M., Haylock, R., Hunter, N. & Zhang, W. Risk of leukemia associated with protracted low-dose radiation exposure: Updated results from the national registry for radiation workers study. Radiat. Res. 192, 527–537 (2019).
doi: 10.1667/RR15358.1
Sugihara, T. et al. Screening of biomarkers for liver adenoma in low-dose-rate γ-ray-irradiated mice. Int. J. Radiat. Biol. 94, 315–326 (2018).
doi: 10.1080/09553002.2018.1439193
Seed, T. M., Fritz, T. E., Tolle, D. V. & Jackson, W. E. Hematopoietic responses under protracted exposures to low daily dose gamma irradiation. Adv. Sp. Res. 30, 945–955 (2002).
doi: 10.1016/S0273-1177(02)00159-X
Waldner, L. et al. The 2019–2020 EURADOS WG10 and RENEB field test of retrospective dosimetry methods in a small-scale incident involving ionizing radiation. Radiat. Res. 195, 253–264 (2021).
Y.C., D. & Hsu, S.-M. Radio-photoluminescence glass dosimeter (RPLGD). In Advances in Cancer Therapy (2011). https://doi.org/10.5772/23710 .
Zorloni, G. et al. Intercomparison of personal and ambient dosimeters in extremely high-dose-rate pulsed photon fields. Radiat. Phys. Chem. 172, 108764 (2020).
doi: 10.1016/j.radphyschem.2020.108764
Musolino, S. V. Absorbed dose determination in external beam radiotherapy: An international code of practice for dosimetry based on standards of absorbed dose to water. Technical Reports Series No. 398. Health Phys. 81, 592–593 (2001).
doi: 10.1097/00004032-200111000-00017
Port, M. et al. Gene expression signature for early prediction of late occurring pancytopenia in irradiated baboons. Ann. Hematol. https://doi.org/10.1007/s00277-017-2952-7 (2017).
doi: 10.1007/s00277-017-2952-7
Port, M. et al. Pre-exposure gene expression in baboons with and without pancytopenia after radiation exposure. Int. J. Mol. Sci. 18, 541 (2017).
doi: 10.3390/ijms18030541
Manning, G. et al. Comparable dose estimates of blinded whole blood samples are obtained independently of culture conditions and analytical approaches. Second RENEB gene expression study. Int. J. Radiat. Biol. 93, 87–98 (2017).
doi: 10.1080/09553002.2016.1227105
O’Brien, G. et al. FDXR is a biomarker of radiation exposure in vivo. Sci. Rep. 8, 1–14 (2018).
doi: 10.1038/s41598-017-19043-w
Britten, R. A. et al. Sleep fragmentation exacerbates executive function impairments induced by protracted low dose rate neutron exposure. Int. J. Radiat. Biol. https://doi.org/10.1080/09553002.2019.1694190 (2019).
doi: 10.1080/09553002.2019.1694190
Farese, A. M. et al. The ability of filgrastim to mitigate mortality following LD50/60 total-body irradiation is administration time-dependent. Health Phys. 106, 39–47 (2014).
doi: 10.1097/HP.0b013e3182a4dd2c
Port, M. et al. Validating baboon ex vivo and in vivo radiation-related gene expression with corresponding human data. Radiat. Res. 189, 389–398 (2018).
doi: 10.1667/RR14958.1
Abend, M. et al. Examining radiation-induced in vivo and in vitro gene expression changes of the peripheral blood in different laboratories for biodosimetry purposes: First RENEB gene expression study. Radiat. Res. 185, 109–123 (2016).
doi: 10.1667/RR14221.1
Bustin, S. A. et al. The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments. Clin. Chem. 55, 611–622 (2009).
doi: 10.1373/clinchem.2008.112797
Dooms, M., Chango, A. & Abdel-Nour, A. L. PCR quantitative (qPCR) et le guide de bonnes pratiques MIQE: Adaptation et pertinence dans le contexte de la biologie clinique. Ann. Biol. Clin. 72, 265–269 (2014).
Kabacik, S., Manning, G., Raffy, C., Bouffler, S. & Badie, C. Time, dose and Ataxia Telangiectasia Mutated (ATM) status dependency of coding and noncoding RNA expression after ionizing radiation exposure. Radiat. Res. 183, 325–337 (2015).
doi: 10.1667/RR13876.1
Manning, G., Kabacik, S., Finnon, P., Bouffler, S. & Badie, C. High and low dose responses of transcriptional biomarkers in ex vivo X-irradiated human blood. Int. J. Radiat. Biol. 89, 512–522 (2013).
doi: 10.3109/09553002.2013.769694
Templin, T. et al. Radiation-induced micro-RNA expression changes in peripheral blood cells of radiotherapy patients. Int. J. Radiat. Oncol. Biol. Phys. 80, 549–557 (2011).
doi: 10.1016/j.ijrobp.2010.12.061
Paul, S. et al. Prediction of in vivo radiation dose status in radiotherapy patients using ex vivo and in vivo gene expression signatures. Radiat. Res. 175, 257–265 (2011).
doi: 10.1667/RR2420.1
Ostheim, P. et al. Identifying a diagnostic window for the use of gene expression profiling to predict acute radiation syndrome. Radiat. Res. 195, 38–46 (2021).
Uehara, Y. et al. Gene expression profiles in mouse liver after long-term low-dose-rate irradiation with gamma rays. Radiat. Res. 174, 611–617 (2010).
doi: 10.1667/RR2195.1
Amundson, S. A. et al. Differential responses of stress genes to low dose-rate γ irradiation. Mol. Cancer Res. 1, 445–452 (2003).
Paul, S., Smilenov, L. B., Elliston, C. D. & Amundson, S. A. Radiation dose-rate effects on gene expression in a mouse biodosimetry model. Radiat. Res. 184, 24–32 (2015).
doi: 10.1667/RR14044.1
Templin, T., Amundson, S. A., Brenner, D. J. & Smilenov, L. B. Whole mouse blood microRNA as biomarkers for exposure to -rays and 56Fe ions. Int. J. Radiat. Biol. 87, 653–662 (2011).
doi: 10.3109/09553002.2010.549537
Macaeva, E., Mysara, M., De Vos, W. H., Baatout, S. & Quintens, R. Gene expression-based biodosimetry for radiological incidents: Assessment of dose and time after radiation exposure. Int. J. Radiat. Biol. 95, 64–75 (2018).
doi: 10.1080/09553002.2018.1511926
Maezawa, H. et al. Survival of mice and hematopoietic stem cells in bone marrow after intermittent total body irradiation. Radiat. Med. Med. Imaging Radiat. Oncol. 5, 215–219 (1987).
Ainsworth, E. J., Afzal, S. M. J., Crouse, D. A., Hanson, W. R. & Fry, R. J. M. Tissue responses to low protracted doses of high LET radiations or photons: Early and late damage relevant to radio-protective countermeasures. Adv. Sp. Res. 9, 299–313 (1989).
doi: 10.1016/0273-1177(89)90453-5
Seed, T. M. et al. Accommodative responses to chronic irradiation: Effects of dose, dose rate, and pharmacological response modifiers. Mil. Med. 167, 82–86 (2002).
doi: 10.1093/milmed/167.suppl_1.82