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Biological Measure of DNA Damage After Single-Ion Microbeam Biological measure of DNA damage after single-ion microbeam irradiation and Monte Carlo simulations Geraldine Gonon, Carmen Villagrasa, Pascale Voisin, Sylvain Meylan, Mohamedamine Benadjaoud, Nicolas Tang, Frank Langner, Hans Rabus, Joan Francesc Barquinero, Ulrich Giesen, et al. To cite this version: Geraldine Gonon, Carmen Villagrasa, Pascale Voisin, Sylvain Meylan, Mohamedamine Benadjaoud, et al.. Biological measure of DNA damage after single-ion microbeam irradiation and Monte Carlo sim- ulations. 16th International Congress of Radiation Research ICRR 2019, Aug 2019, MANCHESTER, France. 2019. hal-02635559 HAL Id: hal-02635559 https://hal.archives-ouvertes.fr/hal-02635559 Submitted on 27 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Copyright N° 404 Biological measure of DNA damage after single-ion microbeam irradiation and Monte Carlo simulations Géraldine Gonon1,Carmen Villagrasa1, Pascale Voisin1, Sylvain Meylan1, Mohamed Amine Benadjaoud1, Nicolas Tang1, Frank Langner2, Hans Rabus2, Joan Francesc Barquinero3, Ulrich Giesen2, Gaëtan Gruel1 1. Institut de Radioprotection et de Sûreté Nucléaire (IRSN), 92260, Fontenay-aux-Roses, France 2. Physikalisch-Technische Bundesanstalt (PTB), Bundesallee 100, 38116 Braunschweig, Germany Contact : [email protected] 3. Universtitat Autònoma de Barcelona, Facultat de Biociències, 08193 Cerdanyola del Vallès, Spain INTRODUCTION Link between track structure and biological effects of ionizing radiation The growing use of hadrons in oncology entails the need to establish more specific dosimetry concepts adapted to their biological efficacy. This Monte Carlo Simulation (GEANT4 DNA) requires more information on the likelihood of subcellular effects, mainly Computes a track for a given ionizing DNA damage, according to how energy is deposited within cells by ionizing particle at nanometer scale particle tracks. Monte Carlo track structure simulation provides a powerful tool for investigating such relation. However, the reliability of simulation Cell nucleus There is a need to produce results can only be assessed by comparison with dedicated biological data. biological data to help set up of the multi-scale simulation What type of biologic data may be relevant and how to produce them? Prediction of DNA damage events BIOLOGICAL DATA Definition of square pattern of irradiation Raw data (53BP1 foci formation) Target: Cultures of primary Human Umbilical Vein Endothelial Cells Cell nucleus 4 µm (HUVEC) with homogeneous DNA density (controlled cytogenetic state and cell cycle stage => G0G1 phase) 1 particle per point Irradiation: Single ion microbeam facility at the PTB allows to control the energy and the number of particles. A pattern of 5 particles per Theoretical coordinates of 5 hits nucleus was chosen (Number of foci per nucleus > Background) Alpha particles Proton DAPI 53BP1 (Tx-Red) 8 MeV (170 keV/µm) 3 MeV (19 keV/µm) 10 MeV (85 keV/µm) 20 MeV (36 keV/µm) Analysis: Statistical evaluation of nuclear foci formation (53BP1 and γ-H2AX) related to a particle traversal was undertaken in a large population of cells. Aim: Estimate the relative frequency of radiation-induced foci formation following a single particle track in cell nuclei 푁푢푚푏푒푟 표푓 푓표푐 Ratio = Merged 푇ℎ푒표푟푒푡푐푎푙 푛푢푚푏푒푟 표푓 푝푎푟푡푐푙푒 ℎ푡푠 γ-H2AX (FITC) BIOLOGICAL DATA ADJUSTED FOR FACTORS LEADING TO EXPERIMENTAL BIAS Adjustment of raw data 푁푢푚푏푒푟 표푓 푓표푐 − 푏푎푐푘푔푟표푢푛푑 푓표푐 1- Foci background 푇ℎ푒표푟푒푡푐푎푙 푛푢푚푏푒푟 표푓 푝푎푟푡푐푙푒 ℎ푡푠 4 µm + + Correction 1+2 +3 2- Estimated number of particle hits reaching each cell nucleus + 4 µm Correction 1+2 Correction 1 푁푢푚푏푒푟 표푓 푓표푐 − 푏푎푐푘푔푟표푢푛푑 푓표푐 + + 퐸푠푡푚푎푡푒푑 푛푢푚푏푒푟 표푓 푝푎푟푡푐푙푒 ℎ푡푠 Biological dimensions and … but the orientation => Irradiation characteristics influence the number of hits per nucleus orientation of the pattern does not in 10-15 % of cell nuclei of each nucleus is specific change 3- Correction due to two particle tracks generating two foci in close proximity to each other, so that it may be analyzed as one focus formation SIMULATION OF PARTICLE INTERACTIONS WITH DNA Corrected biological data vs. Simulation data . Monte Carlo Simulation with Geant4-DNA Simulation of physical and Definition of DNA DSB 2D projection to obtain a number chemical interactions of the taking into account the of simulated foci that would be particle with DNA geometry of DNA model observed by microscopy cell nucleus 29, 2019 29, - Selection criteria of DSB: August 25 August Particle track – • at least 2, 3 or 4 SSBs • on opposite strands of DNA DNA DSB with at least 2 SBs DNA DSB with at least 3 SBs • Separated by 10 bp at most DNA DSB with at least 4 SBs => This is performed thousands of times at different positions of the target in order to take into account the variation of DNA density inside the simulated nucleus. RESULTS AND CONCLUSION Comparable to corrected biological data, the simulated relative frequencies of at least one radiation-induced focus formation per particle track increase gradually. For α particles of LET ranging from 80 to 170 keV·μm-1, a constant DNA damage-induction frequency was found, where 10-30% of the particle traversals did not lead to foci formation. These findings will allow for a better comprehension of the relationship between the topology of energy deposition from particles of different LET and early cell damage. They will also help obtain an accurate estimate of the probability of particle interaction to induce foci formation at DNA damage sites. 16th International Congress of Radiation ResearchRadiation of Congress International 16th This work was carried out within the European Metrology Research Programme (EMRP) Joint Research Project SIB06 “Biologically Weighted Quantities in RadioTherapy” (BioQuaRT). The EMRP was jointly funded by the EMRP participating countries within EURAMET and the The European Union..
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