MHY1485 potentiates immunogenic cell death induction and anti-cancer immunity following irradiation.

Recent in vitro experiments showed that combined treatment with MHY1485, a low-molecular-weight compound, and X-ray irradiation significantly increased apoptosis and senescence in tumor cells, which was associated with oxidative stress, endoplasmic reticulum (ER) stress and p21 stabilization, compared to radiation treatment alone. However, evidence for MHY1485 treatment-mediated suppression of tumor growth in animals is still lacking. Furthermore, it has been shown that ER stress enhances immunogenic cell death (ICD) in tumor cells, as it can exert a favorable influence on the anti-cancer immune system. In the present study, we examined whether co-treatment of MHY1485 and X-ray irradiation induces ICD and in vivo tumor growth suppression using the CT26 and Lewis lung carcinoma murine tumor cell lines. We found that MHY1485 + X-ray treatment promotes ICD more effectively than X-ray treatment alone. MHY1485 suppresses tumor growth in vivo under co-treatment with X-rays and increases INF-γ, tumor necrosis factor, interleukin-2 and interleukin-12 levels in the spleen as well as the presence of CD8+ cells in the tumor. The results suggest that MHY1485 treatment leads to the conversion of irradiated tumors into effective vaccines. Thus, MHY1485 is a promising lead compound for use in combination with radiotherapy.

The Effect of Pre-Operative Verbal Confirmation for Interventional Radiology Physicians on Their Use of Personal Dosimeters and Personal Protective Equipment.

Interventional radiology (IR) physicians must be equipped with personal passive dosimeters and personal protective equipment (PPE); however, they are inconsistently used. Therefore, we aimed to explore practical measures to increase PPE usage and ascertain whether these measures could lead to an actual decrease in exposure doses to IR physicians. Dosimeters and PPE were visually inspected. Then, a pre-operative briefing was conducted as a direct intervention, and the use of dosimeters and PPE was verbally confirmed. Finally, the intervention effect was verified by measuring the use rates and individual exposure doses. Because of the intervention, the use rate markedly improved and was almost 100%. However, both the effective dose rate (effective dose/fluoroscopy time) and the lens equivalent dose rate (lens equivalent dose/fluoroscopy time) showed that the intervention led to a statistically significant increase in exposure (effective dose rate: p = 0.033; lens equivalent dose rate: p = 0.003). In conclusion, the proper use of dosimeters and PPE raised the radiation exposure values for IR physicians immediately after the intervention, which was hypothesized to be due to the inclusion of exposure overlooked to date and the changes in the dosimeter management method from a single- to a double-dosimeter approach.

Changes in Sulfur Metabolism in Mouse Brains following Radon Inhalation.

Therapy using hot springs, including the high-level radioactive gas "radon", is traditionally conducted as an alternative treatment for various diseases. Oxidative-stress-related diseases are inhibited by the enhancement of antioxidative functions following radon inhalation. We have reported that radon inhalation increased the level of anti-oxidants, such as glutathione (G-SH), in the brain and had a protective antioxidative effect against transient global cerebral ischemic injury. However, no studies have yet revealed the changes in G-SH associated substances after radon inhalation. In this study, we comprehensively analyzed several metabolites, focusing on G-SH. Mice were exposed to radon at concentrations of 200, 2000, or 20,000 Bq/m3 for 1, 3, or 10 days. We detected 27 metabolites in the mouse brains. The result showed that the L-methionine levels increased, whereas the levels of urea, glutathione, and sulfite ion decreased under any condition. Although the ratio of G-SH to oxidized glutathione (GS-SG) decreased, glutathione monosulfide (G-S-SH) and cysteine monosulfide (Cys-S-SH) increased after radon inhalation. G-S-SH and Cys-S-SH can produce a biological defense against the imbalance of the redox state at very low-dose irradiation following radon inhalation because they are strong scavengers of reactive oxygen species. Additionally, we performed an overall assessment of high-dimensional data and showed some specific characteristics. We showed the changes in metabolites after radon inhalation using partial least squares-discriminant analysis and self-organizing maps. The results showed the health effects of radon, especially the state of sulfur-related metabolites in mouse brains under the exposure conditions for radon therapy.

Tumor microenvironment regulation - enhanced radio - immunotherapy.

Radiotherapy (RT) is frequently utilized for cancer treatment in clinical practice and has been proved to have immune stimulation potency in recent years. However, its inhibitory effect on tumor growth, especially on tumor metastasis, is still limited by many factors, including the complex tumor microenvironment (TME). Therefore, the TME - regulating SiO2@MnO2 nanoparticles (SM NPs) were prepared and applied to the combination of RT and immunotherapy. In a bilateral animal model, SM NPs not only enhanced the inhibitory effect of RT on primary tumor growth, but also strengthened the abscopal effect to inhibit the growth of distant untreated tumors. As for the distant untreated tumor, 40% of mice showed complete inhibition of tumor growth and 40% showed a suppressed tumor growth. Moreover, SM NPs showed modulation functions for TME through inducing the increase in intracellular levels of oxygen and reactive oxygen species after their reaction with hydrogen peroxide and the main antioxidative agent glutathione in TME. Lastly, SM NPs also effectively induced the increase in the amounts of cytokines secreted by macrophage - like cells, indicating modulation functions for immune responses. This work highlighted a potential strategy of simultaneously inhibiting tumor growth and metastasis through the regulation of TME and immune responses by SM NPs - enhanced radio - immunotherapy.

Monitoring and Protection against Radiation Dose to Eyes of Operators Performing Neuroendovascular Procedures: A Nationwide Study in Japan.

Objective: To meet the new standard of the annual dose limit for the eye lens recommended by the International Commission on Radiation Protection, radiation doses of neuroendovascular procedures in Japanese institutions were investigated. Methods: Radiation doses to operators involved in 304 neuroendovascular procedures at 30 Japanese institutions were prospectively surveyed. The institutions recruited at an annual meeting of the Japanese Society for Neuroendovascular Therapy participated voluntarily. A maximum of 10 wireless dosimeters were attached to the radiation protection (RP) goggles, the ceiling-mounted RP shielding screen, and the operators' forehead and neck over the protective clothing. Doses recorded inside the goggles were defined as eye lens doses for operators who wore RP goggles, while doses to the forehead were defined as eye lens doses for those who did not. The shielding effect rates of the protection devices were calculated, and statistical analysis was performed for the comparison of radiation doses. Results: From 296 analyzed cases, mean eye lens radiation doses per procedure were 0.088 mGy for the left eye and 0.041 mGy for the right eye. For the left eye, that dose without RP equipment was 0.176 mGy and that with RP goggles plus an RP shielding screen was 0.034 mGy. Four parameters, including left eye dose, air kerma at the patient entrance reference point, fluoroscopic time, and the total number of frames, were assessed for five types of neurovascular procedures. Of them, transarterial embolization for dural arteriovenous fistula was associated with the highest eye lens dose at 0.138 mGy. The shielding effect rates of protection goggles were 60% for the left and 55% for the right RP goggle. The mean doses to the inner and outer surfaces of the RP shielding screen were 0.831 mGy and 0.040 mGy, respectively, amounting to a shielding effect rate of 95%. Conclusion: To meet the new standard, both RP goggles and RP shielding screens are strongly recommended to be used effectively. Without proper use of radiological protection devices, the number of neuroendovascular procedures that one operator performs per year will be limited under the new guideline.

Analysis of whole-blood antioxidant capacity after chronic and localized irradiation using the i-STrap method.

Ionizing radiation exposure affects the redox state in vivo. Recently, whole-blood antioxidant capacity (WBAC) has been reported to decrease in a dose-dependent manner after acute total body irradiation (TBI). However, changes in WBAC after localized and chronic irradiations have not been reported. This study analyzed changes to WBAC in mice after either localized irradiation (irradiation of the left hind leg only) or chronic TBI using the i-STrap method. Leg-localized irradiation exerted limited effects on WBAC, while WBAC decreased in a dose rate-dependent manner after TBI. Further, the WBAC reached the minimum value in a shorter period at a smaller dose rate. Our results suggest that changes in WBAC do not directly reflect absorbed dose, but may reflect radiation-induced biological damage at the systemic level. This study will contribute to the understanding of radiation-induced injuries and diseases, and will facilitate the establishment of biomarkers for radiation exposure.

Biosafety of mesoporous silica nanoparticles: a combined experimental and literature study.

Mesoporous silica (MS) particles have been explored for various healthcare applications, but universal data about their safety and/or toxicity are yet to be well-established for clinical purposes. Information about general toxicity of hollow MS (HMS) particles and about immunotoxicity of MS particles are significantly lacked. Therefore, acute toxicity and immunotoxicity of HMS particles were experimentally evaluated. A systematic and objective literature study was parallelly performed to analyze the published in vivo toxicity of MS particles. Lethal acute toxicity of MS particles is likely to arise from their physical action after intravenous and intraperitoneal administrations, and only rarely observed after subcutaneous administration. No clear relationship was identified between physicochemical properties of MS particles and lethality as well as maximum tolerated dose with some exceptions. At sub-lethal doses, MS particles tend to accumulate mainly in lung, liver, and spleen. The HMS particles showed lower inflammation-inducing ability than polyinosinic-polycytidylic acid and almost the same allergy-inducing ability as Alum. Finally, the universal lowest observed adverse effect levels were determined as 0.45, 0.81, and 4.1 mg/kg (human equivalent dose) for intravenous, intraperitoneal, and subcutaneous administration of MS particles, respectively. These results could be helpful for determining an appropriate MS particle dose in clinical study.

MHY1485 enhances X-irradiation-induced apoptosis and senescence in tumor cells.

The mammalian target of rapamycin (mTOR) is a sensor of nutrient status and plays an important role in cell growth and metabolism. Although inhibition of mTOR signaling promotes tumor cell death and several mTOR inhibitors have been used clinically, recent reports have shown that co-treatment with MHY1485, an mTOR activator, enhances the anti-cancer effects of anti-PD-1 antibody and 5-fluorouracil. However, it remains unclear whether MHY1485 treatment alters the effects of radiation on tumor cells. In this study, the radiosensitizing effects of MHY1485 were investigated using murine CT26 and LLC cell lines. We examined mTOR signaling, tumor cell growth, colony formation, apoptosis, senescence, oxidative stress, p21 accumulation and endoplasmic reticulum (ER) stress levels in cells treated with MHY1485 and radiation, either alone or together. We found that MHY1485 treatment inhibited growth and colony formation in both cell lines under irradiation and no-irradiation conditions, results that were not fully consistent with MHY1485's known role in activating mTOR signaling. Furthermore, we found that combined treatment with MHY1485 and radiation significantly increased apoptosis and senescence in tumor cells in association with oxidative stress, ER stress and p21 stabilization, compared to radiation treatment alone. Our results suggested that MHY1485 enhances the radiosensitivity of tumor cells by a mechanism that may differ from MHY1485's role in mTOR activation.

Neurobehavioral effects of acute low-dose whole-body irradiation.

Radiation exposure has multiple effects on the brain, behavior and cognitive functions. It has been reported that high-dose (>20 Gy) radiation-induced behavior and cognitive aberration partly associated with severe tissue destruction. Low-dose (<3 Gy) exposure can occur in radiological disasters and cerebral endovascular treatment. However, only a few reports analyzed behavior and cognitive functions after low-dose irradiation. This study was undertaken to assess the relationship between brain neurochemistry and behavioral disruption in irradiated mice. The irradiated mice (0.5 Gy, 1 Gy and 3 Gy) were tested for alteration in their normal behavior over 10 days. A serotonin (5-HT), Dopamine, gamma-Aminobutyric acid (GABA) and cortisol analysis was carried out in blood, hippocampus, amygdala and whole brain tissue. There was a significant decline in the exploratory activity of mice exposed to 3 Gy and 1 Gy radiation in an open field test. We observed a significant short-term memory loss in 3 Gy and 1 Gy irradiated mice in Y-Maze. Mice exposed to 1 Gy and 3 Gy radiation exhibited increased anxiety in an elevated plus maze (EPM). The increased anxiety and memory loss patterns were also seen in 0.5 Gy irradiated mice, but the results were not statistically significant. In this study we observed that neurotransmitters are significantly altered after irradiation, but the neuronal cells in the hippocampus were not significantly affected. This study suggests that the low-dose radiation-induced cognitive impairment may be associated with the neurochemical in low-dose irradiation and unlike the high-dose scenario might not be directly related to the morphological changes in the brain.

Total body irradiation causes a chronic decrease in antioxidant levels.

Ionizing radiation exposure may not only cause acute radiation syndrome, but also an increased risk of late effects. It has been hypothesized that induction of chronic oxidative stress mediates the late effects of ionizing radiation. However, only a few reports have analyzed changes in long-term antioxidant capacity after irradiation in vivo. Our previous study demonstrated changes in whole-blood antioxidant capacity and red blood cell (RBC) glutathione levels within 50 days after total body irradiation (TBI). In this study, seven-week-old, male, C57BL/6J mice exposed to total body irradiation by X-ray and changes in whole-blood antioxidant capacity and RBC glutathione levels at ≥ 100 days after TBI were investigated. Whole-blood antioxidant capacity was chronically decreased in the 5-Gy group. The RBC reduced glutathione (GSH) level and the GSH/oxidative glutathione (GSSG) ratio were chronically decreased after ≥ 1 Gy of TBI. Interestingly, the complete blood counts (CBC) changed less with 1-Gy exposure, suggesting that GSH and the GSH/GSSG ratio were more sensitive radiation exposure markers than whole-blood antioxidant capacity and CBC counts. It has been reported that GSH depletion is one of the triggers leading to cataracts, hypertension, and atherosclerosis, and these diseases are also known as radiation-induced late effects. The present findings further suggest that chronic antioxidant reduction may contribute to the pathogenesis of late radiation effects.

Optimization of the Maximum Skin Dose Measurement Technique Using Digital Imaging and Communication in Medicine-Radiation Dose Structured Report Data for Patients Undergoing Cerebral Angiography.

Understanding the maximum skin dose is important for avoiding tissue reactions in cerebral angiography. In this study, we devised a method for using digital imaging and communication in medicine-radiation dose structured report (DICOM-RDSR) data to accurately estimate the maximum skin dose from the total air kerma at the patient entrance reference point (Total Ka,r). Using a test data set (n = 50), we defined the mean ratio of the maximum skin dose obtained from measurements with radio-photoluminescence glass dosimeters (RPLGDs) to the Total Ka,r as the conversion factor, CFKa,constant, and compared the accuracy of the estimated maximum skin dose obtained from multiplying Total Ka,r by CFKa,constant (Estimation Model 1) with that of the estimated maximum skin dose obtained from multiplying Total Ka,r by the functional conversion factor CFKa,function (Estimation Model 2). Estimation Model 2, which uses the quadratic function for the ratio of the fluoroscopy Ka,r to the Total Ka,r (Ka,r ratio), provided an estimated maximum skin dose closer to that obtained from direct measurements with RPLGDs than compared with that determined using Estimation Model 1. The same results were obtained for the validation data set (n = 50). It was suggested the quadratic function for the Ka,r ratio provides a more accurate estimate of the maximum skin dose in real time.

Identification of Potential Biomarkers of Radiation Exposure in Blood Cells by Capillary Electrophoresis Time-of-Flight Mass Spectrometry.

Biodosimetry is a useful method for estimating personal exposure doses to ionizing radiation. Studies have identified metabolites in non-cellular biofluids that can be used as markers in biodosimetry. Levels of metabolites in blood cells may reflect health status or environmental stresses differentially. Here, we report changes in the levels of murine blood cell metabolites following exposure to X-rays in vivo. Levels of blood cell metabolites were measured by capillary electrophoresis time-of-flight mass spectrometry. The levels of 100 metabolites were altered substantially following exposure. We identified 2-aminobutyric acid, 2'-deoxycytidine, and choline as potentially useful markers of radiation exposure and established a potential prediction panel of the exposure dose using stepwise regression. Levels of blood cell metabolites may be useful biomarkers in estimating exposure doses during unexpected radiation incidents.

TREATMENT OF INTERNAL CAROTID ANEURYSMS USING PIPELINE EMBOLIZATION DEVICES: MEASURING THE RADIATION DOSE OF THE PATIENT AND DETERMINING THE FACTORS AFFECTING IT.

The purpose of this study was to measure the peak skin dose (PSD) and bilateral lens doses using radiophotoluminescence glass dosimeters and to determine the factors influencing the radiation dose in cases of cerebral aneurysm treated with pipeline embolization devices (PEDs). The cumulative dose, PSD and right and left lens doses were 3818.1 ± 1604.6, 1880.0 ± 723.0, 124.8 ± 49.2 and 180.7 ± 124.8 mGy, respectively. Using multivariate analysis, body mass index (p < 0.01; odds ratio (OR) = 1.806; 95% confidence interval (CI) = 1.007-3.238) and deployment time of PED (p < 0.05; OR = 1.107; 95% CI = 1.001-1.224) were found to be the independent predictors of PSD exceeding 2 Gy. Measures such as collimation of the radiation field and optimization of radiation dose should be taken to reduce the radiation to the patient.

Human RECQL4 represses the RAD52-mediated single-strand annealing pathway after ionizing radiation or cisplatin treatment.

Ionizing radiation (IR) and cisplatin are frequently used cancer treatments, although the mechanisms of error-prone DNA repair-mediated genomic instability after anticancer treatment are not fully clarified yet. RECQL4 mutations mainly in the C-terminal region of the RECQL4 gene lead to the cancer-predisposing Rothmund-Thomson syndrome, but the function of RECQL4ΔC (C-terminus deleted) in error-prone DNA repair remains unclear. We established several RECQL4ΔC cell lines and found that RECQL4ΔC cancer cells, but not RECQL4ΔC nontumorigenic cells, exhibited IR/cisplatin hypersensitivity. Notably, RECQL4ΔC cancer cells presented increased RPA2/RAD52 foci after cancer treatments. RECQL4ΔC HCT116 cells exhibited increased error-prone single-strand annealing (SSA) activity and decreased alternative end-joining activities, suggesting that RECQL4 regulates the DNA repair pathway choice at double-strand breaks. RAD52 depletion by siRNA or RAD52 inhibitors (5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside [AICAR], (-)-epigallocatechin [EGC]) or a RAD52-phenylalanine 79 aptamer significantly restrained the growth of RAD52-upregulated RECQL4ΔC HCT116 cells in vitro and in mouse xenografts. Remarkably, compared to single-agent cisplatin or EGC treatment, cisplatin followed by low-concentration EGC had a significant suppressive effect on RECQL4ΔC HCT116 cell growth in vivo. Together, the regimens targeting the RAD52-mediated SSA pathway after anticancer treatment may be applicable for cancer patients with RECQL4 gene mutations.

Pilot clinical study of ascorbic acid treatment in cardiac catheterization.

Clinical radiodiagnosis and radiotherapy sometimes induce tissue damage and/or increase the risk of cancer in patients. However, in radiodiagnosis, a reduction in the exposure dose causes a blockier image that is not acceptable for diagnosis. Approximately 70% of DNA damage is induced via reactive oxygen species and/or radicals created during X-ray irradiation. Therefore, treatment with anti-oxidants and/or radical scavengers is considered to be effective in achieving a good balance between image quality and damage. However, few studies have examined the effect of using radical scavengers to reduce radiation damage in the clinical setting. In this study, we administrated 20 mg/kg ascorbic acid (AA) to patients before cardiac catheterization (CC) for diagnostic purposes. We analyzed changes in the number of phosphorylated H2AX (γH2AX) foci (a marker of DNA double-strand breaks) in lymphocytes, red blood cell glutathione levels, blood cell counts, and biochemical parameters. Unfortunately, we did not find satisfactory evidence to show that AA treatment reduces γH2AX foci formation immediately after CC. AA treatment did, however, cause a higher reduced/oxidized glutathione ratio than in the control arm immediately after CC. This is a preliminary study, but this result suggests that reducing radiation damage in clinical practice can be achieved using a biological approach.

[The First Step in the Optimization of Radiation Protection of Patients in Cerebral Angiography: Investigate the Possibility of Constructing the Diagnostic Reference Level by Imaging Objective/Disease Group Using Display Value of the Blood Vessel Imaging Apparatus].

To optimize the radiation protection of patients, we investigated the possibility of constructing the diagnostic reference levels (DRLs) by imaging objective/disease group using display value of the blood vessel imaging apparatus (air kerma-area product: PKA, air kerma at the patient entrance reference point: Ka, r) in cerebral angiography. We used PKA and Ka, r recorded during surgery of 997 patients at our hospital, and classified them according to the purpose of imaging (diagnostic cerebral angiography or neuro interventional radiology) and disease group. Neuro interventional radiology (PKA: 268±155 Gy・cm2, Ka, r: 2420±1462 mGy) was significantly higher than that of diagnostic cerebral angiography (PKA: 161±70 Gy・cm2, Ka, r: 1112±485 mGy), (Mann-Whitney test, P<0.01). Significant difference was found between PKA and Ka, r for imaging purpose and disease group (Kruskal-Wallis test, P<0.05). It is highly probable that the DRL for cerebral angiography can be constructed by imaging purpose/disease group using display value (PKA, Ka, r) of the blood vessel imaging apparatus.

ESTIMATION OF PATIENT LENS DOSE ASSOCIATED WITH C-ARM CONE-BEAM COMPUTED TOMOGRAPHY USAGE DURING INTERVENTIONAL NEURORADIOLOGY.

The purpose of this study was to investigate the dose distribution and lens doses associated with C-arm cone-beam computed tomography (CBCT), using a head phantom, and to estimate the contribution ratio of C-arm CBCT to each patient's lens dose during interventional neuroradiology ('lens dose ratio') in 109 clinical cases. In the phantom study, the peak skin doses and respective right and left lens doses of C-arm CBCT were as follows: 63.0 ± 1.9 mGy, 19.7 ± 1.4 mGy and 21.9 ± 0.8 mGy in whole brain C-arm CBCT and 39.2 ± 1.4 mGy, 4.7 ± 0.9 mGy and 3.6 ± 0.3 mGy in high-resolution C-arm CBCT. In the clinical study, the lens dose ratios were 25.4 ± 8.7% in the right lens and 19.1 ± 9.8% in the left lens. This study shows that, on average, ~25% of patients' total lens dose was contributed by C-arm CBCT.

Administration of Dendritic Cells and Anti-PD-1 Antibody Converts X-ray Irradiated Tumors Into Effective In situ Vaccines.

PURPOSE: Danger signals and release of tumor-specific antigens after exposure to ionizing radiation can convert an irradiated tumor into an in situ vaccine. However, radiation alone is not sufficient to induce an effective systemic immune response. In this study, we investigated whether a combination of x-ray irradiation with bone marrow-derived dendritic cells (BM-DCs) and anti-PD-1 antibody (αPD1-ab) administration can enhance both local tumor control and the systemic abscopal effect in murine subcutaneous tumor models. METHODS AND MATERIALS: B16/BL6 melanoma and Lewis lung carcinoma cells were examined for radiosensitivity and expression of H-2kd and PD-L1 before and after irradiation. The tumor cells were implanted subcutaneously in the left thigh of C57BL/6 mice as primary tumors. BM-DCs were induced from mouse bone marrow cells using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). The primary tumors were treated with 8 Gy of x-ray, followed by simultaneous intratumoral injection of BM-DCs and intraperitoneal injection of αPD1-ab. To examine the abscopal effect, the same tumor cells were also inoculated in the right thigh as metastatic tumors 4 days after the primary tumor inoculation, and only the primary tumors were treated with the same protocols. In vivo analyses of tumor growth and survival rates and in vitro analyses of splenic T-cell proliferation and interferon-γ release were performed. RESULTS: The triple-combination treatment of x-ray irradiation with BM-DC and αPD1-ab administration inhibited primary tumor growth and significantly extended survival time in association with significant increase of T-cell proliferation and interferon-γ release. In addition, this triple-combination treatment significantly inhibited the growth of metastatic tumors. CONCLUSIONS: The results indicated that BM-DC and αPD1-ab administration led to the conversion of irradiated tumors into effective in situ vaccines. This combination therapy can be a promising approach to develop a novel individualized therapy for patients with solid cancers.

Dose-dependent decrease in anti-oxidant capacity of whole blood after irradiation: A novel potential marker for biodosimetry.

Many reports have demonstrated that radiation stimulates reactive oxygen species (ROS) production by mitochondria for a few hours to a few days after irradiation. However, these studies were performed using cell lines, and there is a lack of information about redox homeostasis in irradiated animals and humans. Blood redox homeostasis reflects the body condition well and can be used as a diagnostic marker. However, most redox homeostasis studies have focused on plasma or serum, and the anti-oxidant capacity of whole blood has scarcely been investigated. Here, we report changes in the anti-oxidant capacity of whole blood after X-ray irradiation using C57BL/6 J mice. Whole-blood anti-oxidant capacity was measured by electron spin resonance (ESR) spin trapping using a novel spin-trapping agent, 2-diphenylphosphinoyl-2-methyl-3,4-dihydro-2H-pyrrole N-oxide (DPhPMPO). We found that whole-blood anti-oxidant capacity decreased in a dose-dependent manner (correlation factor, r > 0.9; P < 0.05) from 2 to 24 days after irradiation with 0.5-3 Gy. We further found that the red blood cell (RBC) glutathione level decreased and lipid peroxidation level increased in a dose-dependent manner from 2 to 6 days after irradiation. These findings suggest that blood redox state may be a useful biomarker for estimating exposure doses during nuclear and/or radiation accidents.

Metabolic analysis of radioresistant medulloblastoma stem-like clones and potential therapeutic targets.

Medulloblastoma is a fatal brain tumor in children, primarily due to the presence of treatment-resistant medulloblastoma stem cells. The energy metabolic pathway is a potential target of cancer therapy because it is often different between cancer cells and normal cells. However, the metabolic properties of medulloblastoma stem cells, and whether specific metabolic pathways are essential for sustaining their stem cell-like phenotype and radioresistance, remain unclear. We have established radioresistant medulloblastoma stem-like clones (rMSLCs) by irradiation of the human medulloblastoma cell line ONS-76. Here, we assessed reactive oxygen species (ROS) production, mitochondria function, oxygen consumption rate (OCR), energy state, and metabolites of glycolysis and tricarboxylic acid cycle in rMSLCs and parental cells. rMSLCs showed higher lactate production and lower oxygen consumption rate than parental cells. Additionally, rMSLCs had low mitochondria mass, low endogenous ROS production, and existed in a low-energy state. Treatment with the metabolic modifier dichloroacetate (DCA) resulted in mitochondria dysfunction, glycolysis inhibition, elongated mitochondria morphology, and increased ROS production. DCA also increased radiosensitivity by suppression of the DNA repair capacity through nuclear oxidization and accelerated the generation of acetyl CoA to compensate for the lack of ATP. Moreover, treatment with DCA decreased cancer stem cell-like characters (e.g., CD133 positivity and sphere-forming ability) in rMSLCs. Together, our findings provide insights into the specific metabolism of rMSLCs and illuminate potential metabolic targets that might be exploited for therapeutic benefit in medulloblastoma.