¹⁸F-fluoro-deoxyglucose-positron emission tomography (FDG-PET) is a molecular imaging technique that can be used to monitor treatment-related changes in tumor glucose uptake. In cervical cancer, the FDG-PET response is predictive of survival outcome after chemoradiation. In an effort to understand the signaling pathways that regulate the FDG-PET response in cervical cancer, gene expression profiling was performed. Gene set enrichment analysis (GSEA) identified alterations in expression of genes from the PI3K/AKT signaling pathway in tumors with incomplete FDG-PET response after chemoradiation. Pretreatment phosphorylation and activation of AKT is common in cervical cancer, and patients whose tumors have increased pAKT expression prior to treatment have inferior survival outcomes after standard chemoradiation. Recently, activating mutations in the PIK3CA gene have been identified that are associated with poor prognosis after chemoradiation in cervical cancer. These results suggest that targeted inhibition of PI3K/AKT may improve response to chemoradiation in cervical cancer.

The effect of transgenerational exposure to low dose rate (2.4 and 21 mGy/day) gamma irradiation on the yield of DNA double-strand breaks and oxidized guanine (8-hydroxyguanine) has been studied in the muscle and liver tissue of a model organism, the Japanese medaka fish. We found the level of unrepaired 8-hydroxyguanine in muscle tissue increased nonlinearly over four generations and the pattern of this change depended on the radiation dose rate, suggesting that our treatment protocols initiated genomic instability and an adaptive response as the generations progressed. The yield of unrepaired double-strand breaks did not vary significantly among successive generations in muscle tissue in contrast to liver tissue in which it varied in a nonlinear manner. The 8-hydroxyguanine and DSB radiation yields were significantly higher at 2.4 mGy/day than at 21 mGy/day in both muscle and liver tissue in all generations. These data are consistent with the hypothesis of a threshold for radiation-induced activation of DNA repair systems below which tissue levels of DNA repair enzymes remain unchanged, leading to the accumulation of unrepaired damage at very low doses and dose rates.

The objective of this study was to determine whether radiation-induced injury to the heart after 10 Gy total body irradiation (TBI) is direct or indirect. Young male WAG/RijCmcr rats received a 10 Gy single dose using TBI, upper hemi-body (UHB) irradiation, lower hemi-body (LHB) irradiation, TBI with the kidneys shielded or LHB irradiation with the intestines shielded. Age-matched, sham-irradiated rats served as controls. The lipid profile, kidney injury, heart and liver morphology and cardiac function were determined up to 120 days after irradiation. LHB, but not UHB irradiation, increased the risk factors for cardiac disease as well as the occurrence of cardiac and kidney injury in a way that was quantitatively and qualitatively similar to that observed after TBI. Shielding of the kidneys prevented the increases in risk factors for cardiac disease. Shielding of the intestines did not prevent the increases in risk factors for cardiac disease. There was no histological evidence of liver injury 120 days after irradiation. Injury to the heart from irradiation appears to be indirect, supporting the notion that injury to abdominal organs, principally the kidneys, is responsible for the increased risk factors for and the occurrence of cardiac disease after TBI and LHB irradiation.

Methyl-2-cyano-3,12 dioxoolean-1,9 diene-28-oate (CDDO-Me) is an antioxidative, anti-inflammatory modulator, which activates the nuclear factor-erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway. While CDDO-Me has radioprotective activity through Nrf2 activation in vitro and in vivo, its ability to mitigate radiation-induced damage when provided after irradiation has not been studied. Here we investigated whether CDDO-Me mitigates ionizing radiation (IR)-induced DNA damage in immortalized normal human colonic epithelial cells (HCECs) and bronchial epithelial cells (HBECs). DNA damage and clonogenic survival were assessed after treatment with CDDO-Me postirradiation. We observed that treatment with CDDO-Me within 30 min after irradiation improved both DNA damage repair and clonogenic survival independently of Nrf2. CDDO-Me activates the epidermal growth factor receptor (EGFR) related DNA repair responses. In the presence of CDDO-Me, EGFR is phosphorylated and translocates into the nucleus where it interacts with DNA-PKcs. CDDO-Me-mediated mitigation activity can be abrogated through depletion of EGFR, ectopic overexpression of mutant EGFR or inhibition of DNA-PKcs. While post-treatment of CDDO-Me protected noncancerous HCECs and HBECs against IR, cancer cells (HCT116 and MCF7) were not protected by CDDO-Me. These results suggest that targeting EGFR using CDDO-Me is a promising radiation mitigator with potential utility for first responders to nuclear accidents.

Recently, radiation-induced bystander effects (RIBE) have been studied in mouse models in vivo, which clearly demonstrated bystander effects among somatic cells. However, there is currently no evidence for RIBE between somatic cells and germ cells in animal models in vivo. In the current study, the model animal Caenorhabditis elegans was used to investigate the bystander signaling from somatic cells to germ cells, as well as underlying mechanisms. C. elegans body size allows for precise microbeam irradiation and the abundant mutant strains for genetic dissection relative to currently adopted mouse models make it ideal for such analysis. Our results showed that irradiation of posterior pharynx bulbs and tails of C. elegans enhanced the level of germ cell apoptosis in bystander gonads. The irradiation of posterior pharynx bulbs also increased the level of DNA damage in bystander germ cells and genomic instability in the F1 progeny of irradiated worms, suggesting a potential carcinogenic risk in progeny even only somatic cells of parents are exposed to ionizing radiation (IR). It was also shown that DNA damage-induced germ cell death machinery and MAPK signaling pathways were both involved in the induction of germ cell apoptosis by microbeam induced bystander signaling, indicating a complex cooperation among multiple signaling pathways for bystander effects from somatic cells to germ cells.

Events such as a nuclear meltdown accident or nuclear attack have potential for severe radiation injuries. Radiation injury frequently occurs in combination with other forms of trauma, most often burns. Thus far, combined injury studies have focused mainly on skin wound healing and damage to the gut. Since both radiation exposure and remote burn have pulmonary consequences, we examined the early effects of combined injury on the lung. C57BL/6 male mice were irradiated with 5 Gy of total body irradiation followed by a 15% total body surface area scald burn. Lungs from surviving animals were examined for evidence of inflammation and pneumonitis. At 48 h post-injury, pathology of the lungs from combined injury mice showed greater inflammation compared to all other treatment groups, with marked red blood cell and leukocyte congestion of the pulmonary vasculature. There was excessive leukocyte accumulation, primarily neutrophils, in the vasculature and interstitium, with occasional cells in the alveolar space. At 24 and 48 h post-injury, myeloperoxidase levels in lungs of combined injury mice were elevated compared to all other treatment groups (P < 0.01), confirming histological evidence of neutrophil accumulation. Pulmonary levels of the neutrophil chemoattractant KC (CXCL1) were 3 times above that of either injury alone (P < 0.05). Further, monocyte chemotactic protein-1 (MCP-1, CCL2) was increased two- and threefold compared to burn injury or radiation injury, respectively (P < 0.05). Together, these data suggest that combined radiation and burn injury augments early pulmonary congestion and inflammation. Currently, countermeasures for this unique type of injury are extremely limited. Further research is needed to elucidate the mechanisms behind the synergistic effects of combined injury in order to develop appropriate treatments.

The frequency of binucleated cells containing one or more micronuclei (MNBN cells) in cytokinesis-blocked peripheral blood lymphocytes can be used to determine whether a person has received an overexposure to ionizing radiation. However, the absence of a pre-exposure sample can preclude precise dosimetry. Here we use a database of MNBN cell frequencies in peripheral blood lymphocytes from 3,104 apparently healthy, unexposed, control subjects aged birth to 88 years, contributed by laboratories participating in the HUMN project. To determine whether a putatively exposed person has actually received a measurable dose, that person's peripheral blood lymphocyte MNBN frequency is compared to values from age and gender-matched controls in the database. If the subject's frequency is significantly higher than the controls, then a cobalt-60 dose–response curve obtained with the cytokinesis-block micronucleus (CBMN) assay in human peripheral blood lymphocytes is used to estimate the minimum dose of low-LET radiation that could have caused the increase. The response curve was generated with 11 acutely administered doses ranging from 0–4 Gy; the majority of doses were in the low end of this range to provide an accurate estimate of the linear portion of the response. The minimum detectable acute whole-body dose at the 95% prediction level and their corresponding 95% confidence intervals are 0.18 Gy (0.15–0.22) and 0.20 (0.17–0.24) Gy for 20-year-old males and females, respectively. Corresponding values for 50 year olds are 0.23 Gy (0.19–0.26) and 0.25 (0.21–0.29) Gy, and for 70 year olds are 0.24 (0.21–0.28) Gy and 0.26 (0.22–0.31) Gy. The minimum detectable chronic doses are approximately fivefold higher for both genders. These types of analyses, including knowledge of assay variability, will improve our understanding of the requirements and limitations for biodosimetry when a pre-exposure micronucleus value is unavailable and reliance on historical baseline micronucleus values is required.

The ability of radiation to increase the invasiveness of cancer cells is associated with the inflammatory response, which is induced in almost all irradiated patients. For breast cancer patients, elevated plasma levels of the inflammatory cytokine interleukin-1β (IL1β) persisted for a few weeks after completion of radiotherapy. The aim of this study was to determine whether IL1β is involved in the enhancement of breast cancer cell invasion induced by radiation. The role of IL1β was assessed with invasion chambers where irradiated fibroblasts were used as chemoattractant for the MDA-MB-231 breast cancer cells plated in the upper compartment. The ability of IL1β to stimulate the expression of cyclooxygenase-2 (COX-2) and biosynthesis of the prostaglandin E2 (PGE2) in MDA-MB-231 cells were also determined. Our results show that radiation-enhancement of MDA-MB-231 cell invasion was prevented with an anti-IL1β antibody. The production of IL1β was increased in irradiated fibroblasts, while the invasiveness of the MDA-MB-231 cells not exposed to irradiated fibroblasts was favored by adding this cytokine. Furthermore, addition of the COX-2 inhibitor NS-398 prevented the stimulation of cancer cell invasion induced either by irradiated fibroblasts or IL1β. We propose that the effect of IL1β on the invasiveness of the MDA-MB-231 cells involves elevation of matrix metalloproteinase-9 (MMP-9) production, induction of COX-2 expression and PGE2 biosynthesis. In conclusion, this study supports the involvement of IL1β in the radiation-enhancement of breast cancer cell invasion.

After the Tokyo Electric Power Company Fukushima Daiichi nuclear power plant accident on March 11, 2011, the reconstruction of early internal radiation doses in residents of Fukushima plays a major role in evaluating their future heath risk, including thyroid cancer by internal radioiodine. Internal radioactivity was measured using a whole body counter (WBC) at the Nagasaki University Medical School to evaluate the health risks of residents and short term visitors in Fukushima. Measurable ¹³¹I, ¹³⁴Cs and ¹³⁷Cs were detected altogether in 49 out of 196 people who were in Fukushima prefecture at any time during March 11 and April 20, 2011. In 49 people, the 90 percentile of the thyroid equivalent dose by ¹³¹I and the committed effective dose (total effective dose over a lifetime) by the sum of ¹³⁴Cs and ¹³⁷Cs was 3 mSv and 0.06 mSv, respectively. The radionuclide intakes in early evacuees who left Fukushima before March 16 were more than five times as high as in the responders who moved to Fukushima later. The intake ratio of ¹³¹I/¹³⁷Cs of the earlier evacuees was approximately three. The spatial analysis of 16 evacuees to the south indicated a reduction of internal radioactivity depending on the distance from the nuclear power plant. Among them, high internal ¹³¹I radioactivity in 6 people in a particular evacuation route could be explained by the arrival of a radioactive cloud with a high airborne ¹³¹I/¹³⁷Cs ratio to the environment, as predicted by atmospheric dispersion simulations. Overall, the actual internal radioactivity assessed by a WBC examination comparatively agreed with the predicted airborne radioactivity. These results suggest that the accurate estimation of internal doses in the first week after the radiological accident is critical for the dose reconstruction. The evaluation of internal doses of residents based on their evacuation routes and the advanced estimation of airborne radioactivity from the atmospheric dispersion model should continue to be assessed.

A mechanism-based, two-parameter biophysical model of cell killing was developed with the aim of elucidating the mechanisms underlying radiation-induced cell death and predicting cell killing by different radiation types, including protons and carbon ions at energies and doses of interest for cancer therapy. The model assumed that certain chromosome aberrations (dicentrics, rings and large deletions, called “lethal aberrations”) lead to clonogenic inactivation, and that aberrations derive from μm-scale misrejoining of chromatin fragments, which in turn are produced by “dirty” double-strand breaks called “cluster lesions” (CLs). The average numbers of CLs per Gy per cell were left as a semi-free parameter and the threshold distance for chromatin-fragment rejoining was defined the second parameter. The model was “translated” into Monte Carlo code and provided simulated survival curves, which were compared with survival data on V79 cells exposed to protons, carbon ions and X rays. The agreement was good between simulations and survival data and supported the assumptions of the model at least for doses up to a few Gy. Dicentrics, rings and large deletions were found to be lethal not only for AG1522 cells exposed to X rays, as already reported by others, but also for V79 cells exposed to protons and carbon ions of different energies. Furthermore, the derived CL yields suggest that the critical DNA lesions leading to clonogenic inactivation are more complex than “clean” DSBs. After initial validation, the model was applied to characterize the particle and LET dependence of proton and carbon cell killing. Consistent with the proton data, the predicted fraction of inactivated cells after 2 Gy protons was 40–50% below 7.7 keV/μm, increased by a factor ∼1.6 between 7.7–30.5 keV/μm, and decreased by a factor ∼1.1 between 30.5–34.6 keV/μm. These LET values correspond to proton energies below a few MeV, which are always present in the distal region of hadron therapy spread-out Bragg peaks (SOBP). Consistent with the carbon data, the predicted fraction of inactivated cells after 2 Gy carbon was 40–50% between 13.7–32.4 keV/μm, it increased by a factor ∼1.7 between 32.4–153.5 keV/μm, and decreased by a factor ∼1.1 between 153.5–339.1 keV/μm. Finally, we applied the model to predict cell death at different depths along a carbon SOBP used for preclinical experiments at HIMAC in Chiba, Japan. The predicted fraction of inactivated cells was found to be roughly constant (less than 10%) along the SOBP, suggesting that this approach may be applied to predict cell killing of therapeutic carbon beams and that, more generally, dicentrics, rings and deletions at the first mitosis may be regarded as a biological dose for these beams. This study advanced our understanding of the mechanisms of radiation-induced cell death and characterized the particle and LET dependence of proton and carbon cell killing along a carbon SOBP. The model does not use RBE values, which can be a source of uncertainty. More generally, this model is a mechanism-based tool that in minutes can predict cell inactivation by protons or carbon ions of a given energy and dose, based on an experimental photon curve and in principle, a single (experimental) survival point for the considered ion type and energy.

Previous studies demonstrated that genistein protects mice from radiation-induced bone marrow failure. To overcome genistein's extremely low water solubility, a nanoparticle suspension of genistein has been formulated for more rapid dissolution. In the current study, we evaluated the radioprotective effects of a nanoparticle formulation of genistein on survival and hematopoietic recovery in mice exposed to total-body gamma irradiation. A single intramuscular injection of a saline-based genistein nanosuspension (150 mg/kg) administered to CD2F1 mice 24 h before 9.25 Gy ⁶⁰Co radiation exposure resulted in a 30-day survival rate of 95% compared to 25% in vehicle-treated animals. In mice irradiated at 7 Gy, the genistein nanosuspension increased mouse bone marrow cellularity from approximately 2.9% (vehicle treated) to 28.3% on day 7 postirradiation. Flow cytometry analysis demonstrated decreased radiation-induced hematopoietic stem and progenitor cell (HSPC, Lineage^–/cKit ) death from 77.0% (vehicle) to 43.9% (genistein nanosuspension) with a significant recovery of clonogenicity 7 days after irradiation. The genistein nanosuspension also attenuated the radiation-induced elevation of proinflammatory factors interleukin 1 beta (IL-1β), IL-6 and cyclooxygenase-2 (COX-2) in mouse bone marrow and spleen, which may contribute to protecting HSPCs.

Cranial irradiation is a critical and effective treatment for primary brain tumors and metastases. Unfortunately, most patients who are treated and survive for more than a few months develop neural and cognitive problems as the result of radiation-induced normal tissue injury. The neurobiological mechanisms underlying these cognitive deficits remain largely unknown and there are no validated treatments to prevent or ameliorate them; thus, there is a significant and continuing need for preclinical studies in animal models. Investigations from several laboratories have demonstrated neurobiological changes after cranial irradiation in rodents. To date, however, experimental studies in animal models have included little assessment of the systemic effects of cranial irradiation, despite evidence from the clinic that cranial irradiation results in changes throughout the body and recognition that systemic responses may influence the development of neural and cognitive deficits. This study evaluated systemic effects of clinically relevant, fractionated whole-brain irradiation in adult rats and demonstrates effects on the growth hormone/insulin-like growth factor-I axis, which may contribute to the development of neural changes. These and other systemic responses are important to consider in ongoing efforts to understand the mechanisms of radiation-induced normal tissue injury.