RENEB Inter-Laboratory Comparison 2021: The Dicentric Chromosome Assay.

After large-scale radiation accidents where many individuals are suspected to be exposed to ionizing radiation, biological and physical retrospective dosimetry assays are important tools to aid clinical decision making by categorizing individuals into unexposed/minimally, moderately or highly exposed groups. Quality-controlled inter-laboratory comparisons of simulated accident scenarios are regularly performed in the frame of the European legal association RENEB (Running the European Network of Biological and Physical retrospective Dosimetry) to optimize international networking and emergency readiness in case of large-scale radiation events. In total 33 laboratories from 22 countries around the world participated in the current RENEB inter-laboratory comparison 2021 for the dicentric chromosome assay. Blood was irradiated in vitro with X rays (240 kVp, 13 mA, ∼75 keV, 1 Gy/min) to simulate an acute, homogeneous whole-body exposure. Three blood samples (no. 1: 0 Gy, no. 2: 1.2 Gy, no. 3: 3.5 Gy) were sent to each participant and the task was to culture samples, to prepare slides and to assess radiation doses based on the observed dicentric yields from 50 manually or 150 semi-automatically scored metaphases (triage mode scoring). Approximately two-thirds of the participants applied calibration curves from irradiations with γ rays and about 1/3 from irradiations with X rays with varying energies. The categorization of the samples in clinically relevant groups corresponding to individuals that were unexposed/minimally (0-1 Gy), moderately (1-2 Gy) or highly exposed (>2 Gy) was successfully performed by all participants for sample no. 1 and no. 3 and by ≥74% for sample no. 2. However, while most participants estimated a dose of exactly 0 Gy for the sham-irradiated sample, the precise dose estimates of the samples irradiated with doses >0 Gy were systematically higher than the corresponding reference doses and showed a median deviation of 0.5 Gy (sample no. 2) and 0.95 Gy (sample no. 3) for manual scoring. By converting doses estimated based on γ-ray calibration curves to X-ray doses of a comparable mean photon energy as used in this exercise, the median deviation decreased to 0.27 Gy (sample no. 2) and 0.6 Gy (sample no. 3). The main aim of biological dosimetry in the case of a large-scale event is the categorization of individuals into clinically relevant groups, to aid clinical decision making. This task was successfully performed by all participants for the 0 Gy and 3.5 Gy samples and by 74% (manual scoring) and 80% (semiautomatic scoring) for the 1.2 Gy sample. Due to the accuracy of the dicentric chromosome assay and the high number of participating laboratories, a systematic shift of the dose estimates could be revealed. Differences in radiation quality (X ray vs. γ ray) between the test samples and the applied dose effect curves can partly explain the systematic shift. There might be several additional reasons for the observed bias (e.g., donor effects, transport, experimental conditions or the irradiation setup) and the analysis of these reasons provides great opportunities for future research. The participation of laboratories from countries around the world gave the opportunity to compare the results on an international level.

CYTOGENETIC DAMAGES IN LUNG CANCER PATIENTS TREATED BY EXTERNAL RADIATION THERAPY. / TsYTOGENETYChNI POShKODZhENNIa U KhVORYKh NA RAK LEGENI PRY DYSTANTsIY̆NIY̆ PROMENEVIY̆ TERAPIÏ.

OBJECTIVE: to estimate the chromosome type aberrations outcome in lung cancer patients during radiation therapy course depending on radiation energy. MATERIALS AND METHODS: Chromosome aberrations were studied in 16 lung cancer patients examined before treat- ment in the middle and at the end of external gamma-radiotherapy 60Со on ROCUS-AM and megavolt therapy on lin- ear accelerator Clinac 600C. RESULTS: The radiation-induced chromosome aberrations outcome in lung cancer patients during radiotherapy with different irradiation sources was studied. The over-spontaneous excess of chromosome type aberrations in cancer patients before treatment was shown. The similar and different features of cytogenetic damage accumulation dur- ing radiation therapy regarding to radiation type was displayed. The cytogenetic damage frequency raised during the course of therapeutic exposure in both studied groups. At the same time, the growth rate depended on the irra- diation regimen and was higher for patients undergoing 60Co gamma-therapy. The different range of cells with unsta- ble chromosome aberrations from the beginning to the end of the radiation therapy course was displayed. In the middle of the course the number of aberrations per aberrant cell was similar - 1-5 damages for gamma therapy, 1-4 damages for megavoltage therapy. At the end of treatment the maximum of 10 aberrations per aberrant cell for ROCUS-AM and in 5 aberrations per aberrant cell for linear accelerator was observed. During radiotherapy the distri- butions of radiation-induced chromosome aberrations was found to be over-dispersed according to Poisson statis- tic in both patients' groups. CONCLUSIONS: The study of chromosome type aberration revealed the particularities of cytogenetic damages out- come during radiotherapy course depending on the irradiation source used. The more pronounced genotoxic effect in blood lymphocytes of lung cancer patients due to gamma-irradiation with ROCUS-AM was demonstra- ted despite the higher radiation energy used on a linear accelerator. Therefore, in radiation treatment effects estimation it is necessary to take into account not only the radiation quality and energy, but also the source, regimen and pre-radiotherapy procedures. The data obtained can contribute to the assessment of the conse- quences of local fractionated irradiation and to the development of a reference biodosimetry system in radia- tion therapy.

Possibilities and limitations of fluorescence in situ hybridization technique in retrospective detection of low dose radiation exposure in post-chernobyl human cohorts.

Cytogenetic analysis using the fluorescence in situ hybridisation (FISH) technique was performed late time after the Chernobyl accident in groups of liquidators, evacuees from 30 km exclusive zone, residents of radioactively contaminated areas and control donors age-matched to exposed persons. Stable and unstable chromosome type exchanges were recorded using a hybrid conventional-PAINT nomenclature. The mean yield of stable chromosome exchanges in liquidators did not correlate with registered radiation doses but had a clear negative dependence on the duration of liquidators' staying in Chernobyl zone, that was in a good agreement with early data based on conventional dicentrics plus rings analysis. The overspontaneous excess for stable chromosome exchange level appeared to be higher in evacuees 16-40 years old than that of senior persons, whereas no age-dependent difference occurred for initially induced dicentrics plus rings yields in this cohort. The stable chromosome exchange yield, as well as combined yield of dicentrics plus rings and potentially unstable incomplete translocations in residents of radioactively contaminated areas showed a reasonable positive correlation with levels of 137Cs contamination. The observed yields of stable chromosome exchanges in all three exposed groups appeared to be somewhat lower than those of expected from unstable exchange-based doses which were referred to an in vitro dose response of stable exchanges outcome in human lymphocytes. Thus, FISH analysis can be successfully applied for qualitative cytogenetic indication of past and chronic radiation exposure to low doses but further refinement of FISH-based system for quantitative dose assessment is still required. Some practical approaches of solving this task are discussed.

The capacity, capabilities and needs of the WHO BioDoseNet member laboratories.

Biodosimetry is an essential tool for providing timely assessments of radiation exposure, particularly when physical dosimetry is unavailable or unreliable. For mass-casualty events involving public exposure to ionising radiation, it is paramount to rapidly provide this dose information for medical management of casualties. The dicentric chromosome assay is currently the most reliable accepted method for biodosimetry; however, in a mass-casualty scenario, the throughput of this assay will be challenged by its time-consuming nature and the specific expertise required. To address this limitation, many countries have established expertise in cytogenetic biodosimetry and started developing surge capabilities through setting up regional networks to deal with emergency situations. To capitalise on this growing expertise and organise it into an internationally coordinated laboratory network, the World Health Organization has created and launched a global biodosimetry network (BioDoseNet). In order to determine the existing capacity of BioDoseNet member laboratories, including their expertise and in vivo experience, involvement in national and international activities, problems, needs and prospects, an in-depth survey was conducted. These survey results provide significant information on the current state of emergency cytogenetic biodosimetry capabilities around the world.