A simulation approach to predict the calibration curve of low LET radiations for biological dosimetry using the Geant4-DNA toolkit

Ionizing radiation is extensively utilized in various applications; however, it can lead to significant harm to living systems. In this regard, the radiation absorbed dose is usually evaluated by performing biological dosimetry and physical reconstruction of exposure scenarios. But, this is costly, time-consuming, and maybe impractical for a biodosimetry lab to perform biological dosimetry. This study aimed to assess the applicability and reliability of the Geant4-DNA toolkit as a simulation approach to construct a reliable dose-response curve for biodosimetry purposes as an appropriate substitution for experimental measurements. In this matter, the total number of double-strand breaks (DSBs), due to different doses of low LET radiation qualities on DNA molecules, was calculated and converted to the values of dicentric chromosomes using a mechanistic model of cellular response. Then, the number of dicentric chromosomes induced by 200 kVp X-rays were modified by using a semi-empirical scaling factor for compensating the restriction of simulation code to consider what can happen in a real cell. Next, the trend of dicentrics for 137Cs and 60Co were calculated and modified by the above scaling factor. Finally, the dose-response curves for these gamma sources compared to several published experiments. The suggested calibration curves for 137Cs and 60Co followed a linear quadratic equation: Ydic = 0.0054 (± 0.0133) - 0.0089 (± 0.0212) × D + 0.0568 (± 0.0051) × D2 and Ydic = 0.0052 (± 0.0128) - 0.00568 (± 0.0203) × D + 0.0525 (± 0.0049) × D2 respectively. They revealed a satisfactory agreement with the experimental data reported by others. The Geant4 program developed in this work could provide an appropriate tool for predicting the dose-response (calibration) curve for biodosimetry purposes.

Linear Dose Response of Acrocentric Chromosome Associations (Aca) To Gamma Irradiation In Human Lymphocytes

Purpose: The frequency of acrocentric chromosome associations (ACA) was studied to determine the possible dose-response relation with low doses of gamma irradiation in lymphocytes. Methods: Peripheral blood collected from three healthy donors were irradiated with 0, 0.1, 0.25, 0.5, 0.75, and 1 Gy gamma radiation. Chromosomal preparations were made after 48 hrs culture as per the standard guidelines. Results: The average number of ACA and ACA % were increased significantly with an increase in a dose. The D-G and D-D type of association was most prominent and showed a dose-dependent increase. The ACA frequency in irradiated lymphocytes showed an increase concerning the dose. The fitted regression equation was y=0.4759x+0.1663 (R2=0.9635; p=0.0005). An assessment of dicentric chromosomes (DC) was carried for the same slides. The correlation curve was prepared for ACA frequencies versus DC frequencies, resulting in a regression equation as y=8.659x+0.2.37 (R2=0.8275; p=0.0119). Conclusion: Our results showed an increase in frequencies of ACA in irradiated lymphocytes with an increase in radiation dose and followed a similar linear trend with DC frequency, thus, ACA may serve as a candidate cytogenetic biomarker for radiation biodosimetry especially for low radiation doses.

Could scoring tailed and dumbbell-shaped nuclei increase the sensitivity of micronucleus analysis as a biomarker of radiation exposure?

In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear budding (NBUD) are the most commonly analysed morphological types of nuclear abnormalities. In contrast, tailed and dumbbell-shaped nucleus have historically received little attention in the CBMN assay. Interestingly, the incidence of tailed and dumbbell-shaped nuclei in lymphocytes is closely related with that of dicentric chromosomes or NPBs in the CBMN assay. To provide a better picture of the implications and significance of tailed and dumbbell-shaped nuclei as markers of radiation exposure, a literature review was performed in this study. Twenty articles were found in PubMed, PubMed Central, and manually searched. The articles were screened and those that met the inclusion criteria and did not meet the exclusion criteria were reviewed by all authors. At the end, nine articles were included. In conclusion, the assessment of in vivo tailed nuclei in blood smears and accounting for the occurrence of dumbbell-shaped nuclei in the CBMN assay can increase the sensitivity of the CBMN assay for biodosimetry involving a high dose exposure.

Is Response to Genotoxic Stress Similar in Populations of African and European Ancestry? A Study of Dose-Response After in vitro Irradiation

Background: Exposure to genotoxic stress such as radiation is an important public health issue affecting a large population. The necessity of analyzing cytogenetic effects of such exposure is related to the need to estimate the associated risk. Cytogenetic biological dosimetry is based on the relationship between the absorbed dose and the frequency of scored chromosomal aberrations. The influence of confounding factors on radiation response is a topical issue. The role of ethnicity is unclear. Here, we compared the dose-response curves obtained after irradiation of circulating lymphocytes from healthy donors of African and European ancestry.Materials and Methods: Blood samples from six Africans living in Africa, five Africans living in Europe, and five Caucasians living in Europe were exposed to various doses (0–4 Gy) of X-rays at a dose-rate of 0.1 Gy/min using an X-RAD320 irradiator. A validated cohort composed of 14 healthy Africans living in three African countries was included and blood samples were irradiated using the same protocols. Blood lymphocytes were cultured for 48 h and chromosomal aberrations scored during the first mitosis by telomere and centromere staining. The distribution of dicentric chromosomes was determined and the Kruskal-Wallis test was used to compare the dose-response curves of the two populations.Results: No spontaneous dicentric chromosomes were detected in African donors, thus establishing a very low background of unstable chromosomal aberrations relative to the European population. There was a significant difference in the dose response curves between native African and European donors. At 4 Gy, African donors showed a significantly lower frequency of dicentric chromosomes (p = 8.65 10–17), centric rings (p = 4.0310–14), and resulting double-strand-breaks (DSB) (p = 1.32 10–18) than European donors. In addition, a significant difference was found between African donors living in Europe and Africans living in Africa.Conclusion: This is the first study to demonstrate the important role of ethnic and environmental factors that may epigenetically influence the response to irradiation. It will be necessary to establish country-of-origen-specific dose response curves to practice precise and adequate biological dosimetry. This work opens new perspective for the comparison of treatments based on genotoxic agents, such as irradiation.

Cytogenetic bio-dosimetry techniques in the detection of dicentric chromosomes induced by ionizing radiation: A review

Ionizing radiation is ubiquitous in the environment. Its source can be natural, such as radioactive materials present in soil and cosmic rays, or artificial, such as the fuel for nuclear power plants. Overexposure to ionizing radiation may damage living tissue and could cause severe health problems (i.e., mutations, radiation sickness, cancer, and death). Cytogenetic bio-dosimetry has the great advantage to take into account the inter-individual variation, and it is informative even when physical dosimetry is not applicable; moreover, it is the definitive method to assess exposure to ionizing radiation recommended by the World Health Organization (WHO). Such a procedure involves counting the frequency of dicentric chromosomes (DCs), which are the most studied chromosomal aberrations used as absorbed radiation biomarkers, during the metaphase of cells. A set of algorithms, tested on different programming languages to automatically identify DCs, is analyzed by the authors together with an Automated Dicentric Chromosome Identifying software (ADCI) mostly based on OpenCV programming libraries. The purpose of this work is to review the main results regarding the correlation between ionizing radiation and dicentric chromosomes in cytogenetic bio-dosimetry.

Validation of a biomarker tool capable of measuring the absorbed dose soon after exposure to ionizing radiation

A radiological or nuclear attack could involve such a large number of subjects as to overwhelm the emergency facilities in charge. Resources should therefore be focused on those subjects needing immediate medical attention and care. In such a scenario, for the triage management by first responders, it is necessary to count on efficient biological dosimetry tools capable of early detection of the absorbed dose. At present the validated assays for measuring the absorbed dose are dicentric chromosomes and micronuclei counts, which require more than 2–3 days to obtain results. To overcome this limitation the NATO SPS Programme funded an Italian–Egyptian collaborative project aimed at validating a fast, accurate and feasible tool for assessing the absorbed dose early after radiation exposure. Biomarkers as complete blood cell counts, DNA breaks and radio-inducible proteins were investigated on blood samples collected before and 3 h after the first fraction of radiotherapy in patients treated in specific target areas with doses/fraction of about: 2, 3.5 or > 5 Gy and compared with the reference micronuclei count. Based on univariate and multivariate multiple linear regression correlation, our results identify five early biomarkers potentially useful for detecting the extent of the absorbed dose 3 h after the exposure.

Origins of cancer genome complexity revealed by haplotype-resolved genomic analysis of evolution of Barretts esophagus to esophageal adenocarcinoma

Complex chromosomal alterations are a hallmark of advanced cancers but rarely seen in normal tissue. The progression of precancerous lesions to malignancy is often accompanied by increasing complexity of chromosomal alterations that can drive their transformation through focal oncogenic amplifications. However, the etiology and evolution dynamics of these alterations are poorly understood. Here we study chromosomal copy-number evolution in the progression of Barretts esophagus (BE) to esophageal adenocarcinoma (EAC) by multi-regional whole-genome sequencing analysis of BE samples with dysplasia and microscopic EAC foci. Through haplotype-specific copy-number analysis of BE genome evolution, we identified distinct patterns of episodic copy-number evolution consistent with the outcomes of abnormal mitosis and dicentric chromosome breakage. While abnormal mitosis, including whole-genome duplication, accounts for most chromosome or arm-level copy-number changes, segmental copy-number alterations display signatures of multi-generational evolution of unstable dicentric chromosomes. Continuous evolution of dicentric chromosomes through breakage-fusion-bridge cycles and chromothripsis rapidly increases genomic complexity and diversity among BE cells, culminating in the generation of distinct focal amplifications. These mutational processes enable multiple subclones within small dysplastic areas to undergo parallel transformation to cancer following acquisition of distinct oncogenic amplifications. Our results demonstrate how chromosomal instability drives clonal diversification in precancer evolution and promotes tumorigenesis in primary human samples.

Behavior of dicentric chromosomes in budding yeast

DNA double-strand breaks arisein vivowhen a dicentric chromosome (two centromeres on one chromosome) goes through mitosis with the two centromeres attached to opposite spindle pole bodies. Repair of the DSBs generates phenotypic diversity due to the range of monocentric derivative chromosomes that arise. To explore whether DSBs may be differentially repaired as a function of their spatial position in the chromosome, we have examined the structure of monocentric derivative chromosomes from cells containing a suite of dicentric chromosomes in which the distance between the two centromeres ranges from 6.5 kb to 57.7 kb. Two major classes of repair products, homology-based (homologous recombination (HR) and single-strand annealing (SSA)) and end-joining (non-homologous (NHEJ) and micro-homology mediated (MMEJ)) were identified. The distribution of repair products varies as a function of distance between the two centromeres. Genetic dependencies on double strand break repair (Rad52), DNA ligase (Lif1), and S phase checkpoint (Mrc1) are indicative of distinct repair pathway choices for DNA breaks in the pericentromeric chromatin versus the arms.