Grainne Manning1, Ellina Macaeva2,3, Matthaeus Majewski4, Ralf Kriehuber5, Kamil Brzóska6, Michael Abend4, Sven Doucha-Senf4, Dominik Oskamp5, Sonja Strunz7, Roel Quintens2, Matthias Port2, Christophe Badie1. 1. a Cancer Mechanisms and Biomarkers Group, Radiation Effects Department , Centre for Radiation, Chemical and Environmental Hazards, Public Health England , Chilton , UK. 2. b Radiobiology Unit , Institute Environment, Health and Safety, Belgian Nuclear Research Centre , Mol , Belgium. 3. c Department of Molecular Biotechnology , Ghent University , Ghent , Belgium. 4. d Bundeswehr Institute of Radiobiology , Munich , Germany. 5. e Radiation Biology Unit, Department of Safety and Radiation Protection , Forschungszentrum Jülich GmbH, Germany. 6. f Institute of Nuclear Chemistry and Technology , Centre for Radiobiology and Biological Dosimetry , Warsaw , Poland. 7. g Institute of Genetics and Biometry, Leibniz Institute for Farm Animal Biology (FBN) , Dummerstorf , Germany.
Abstract
PURPOSE: This collaboration of five established European gene expression labs investigated the potential impact of culture conditions on the transcriptional response of peripheral blood to radiation exposure. MATERIALS AND METHODS: Blood from one healthy donor was exposed ex vivo to a Cobalt 60 source to produce a calibration curve in addition to four unknown doses. After exposure, the blood samples were either diluted with RPMI medium or left untouched. After 24-h incubation at 37 °C the diluted blood samples were lysed, while the undiluted samples were mixed with the preservative RNALater and all samples were shipped frozen to the participating labs. Samples were processed by each lab using microarray (one lab) and QRT-PCR (four labs). RESULTS: We show that although culture conditions affect the total amount of RNA recovered (p < .0001) and its integrity (p < .0001), it does not significantly affect dose estimates (except for the true dose at 1.1 Gy). Most importantly, the different analysis approaches provide comparable mean absolute difference of estimated doses relative to the true doses (p = .9) and number of out of range (>0.5 Gy) measurements (p = .6). CONCLUSION: This study confirms the robustness of gene expression as a method for biological dosimetry.
PURPOSE: This collaboration of five established European gene expression labs investigated the potential impact of culture conditions on the transcriptional response of peripheral blood to radiation exposure. MATERIALS AND METHODS: Blood from one healthy donor was exposed ex vivo to a Cobalt 60 source to produce a calibration curve in addition to four unknown doses. After exposure, the blood samples were either diluted with RPMI medium or left untouched. After 24-h incubation at 37 °C the diluted blood samples were lysed, while the undiluted samples were mixed with the preservative RNALater and all samples were shipped frozen to the participating labs. Samples were processed by each lab using microarray (one lab) and QRT-PCR (four labs). RESULTS: We show that although culture conditions affect the total amount of RNA recovered (p < .0001) and its integrity (p < .0001), it does not significantly affect dose estimates (except for the true dose at 1.1 Gy). Most importantly, the different analysis approaches provide comparable mean absolute difference of estimated doses relative to the true doses (p = .9) and number of out of range (>0.5 Gy) measurements (p = .6). CONCLUSION: This study confirms the robustness of gene expression as a method for biological dosimetry.
Authors: Lourdes Cruz-Garcia; Grainne O'Brien; Ellen Donovan; Lone Gothard; Sue Boyle; Antoine Laval; Isabelle Testard; Lucyna Ponge; Grzegorz Woźniak; Leszek Miszczyk; Serge M Candéias; Elizabeth Ainsbury; Piotr Widlak; Navita Somaiah; Christophe Badie Journal: Health Phys Date: 2018-07 Impact factor: 1.316
Authors: Lourdes Cruz-Garcia; Grainne O'Brien; Botond Sipos; Simon Mayes; Michael I Love; Daniel J Turner; Christophe Badie Journal: Radiat Res Date: 2019-12-12 Impact factor: 2.841
Authors: M Abend; S A Amundson; C Badie; K Brzoska; R Hargitai; R Kriehuber; G O'Brien; S Schüle; E Kis; S A Ghandhi; K Lumniczky; S R Morton; D Oskamp; P Ostheim; C Siebenwirth; I Shuryak; T Szatmári; M Unverricht-Yeboah; E Ainsbury; C Bassinet; U Kulka; U Oestreicher; Y Ristic; F Trompier; A Wojcik; L Waldner; M Port Journal: Sci Rep Date: 2021-05-07 Impact factor: 4.379
Authors: Ales Tichy; Sylwia Kabacik; Grainne O'Brien; Jaroslav Pejchal; Zuzana Sinkorova; Adela Kmochova; Igor Sirak; Andrea Malkova; Caterina Gomila Beltran; Juan Ramon Gonzalez; Jakub Grepl; Matthaeus Majewski; Elizabeth Ainsbury; Lenka Zarybnicka; Jana Vachelova; Alzbeta Zavrelova; Marie Davidkova; Marketa Markova Stastna; Michael Abend; Eileen Pernot; Elisabeth Cardis; Christophe Badie Journal: PLoS One Date: 2018-02-23 Impact factor: 3.240