Impact of High-Dose Irradiation on Human iPSC-Derived Cardiomyocytes Using Multi-Electrode Arrays: Implications for the Antiarrhythmic Effects of Cardiac Radioablation

Cardiac radioablation is emerging as an alternative option for refractory ventricular arrhythmias. However, the immediate acute effect of high-dose irradiation on human cardiomyocytes remains poorly known. We measured the electrical activities of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) upon irradiation with 0, 20, 25, 30, 40, and 50 Gy using a multi-electrode array, and cardiomyocyte function gene levels were evaluated. iPSC-CMs showed to recover their electrophysiological activities (total active electrode, spike amplitude and slope, and corrected field potential duration) within 3–6 h from the acute effects of high-dose irradiation. The beat rate immediately increased until 3 h after irradiation, but it steadily decreased afterward. Conduction velocity slowed in cells irradiated with ≥25 Gy until 6–12 h and recovered within 24 h; notably, 20 and 25 Gy-treated groups showed subsequent continuous increase. At day 7 post-irradiation, except for cTnT, cardiomyocyte function gene levels increased with increasing irradiation dose, but uniquely peaked at 25–30 Gy. Altogether, high-dose irradiation immediately and reversibly modifies the electrical conduction of cardiomyocytes. Thus, compensatory mechanisms at the cellular level may be activated after the high-dose irradiation acute effects, thereby, contributing to the immediate antiarrhythmic outcome of cardiac radioablation for refractory ventricular arrhythmias.

ROS- and Radiation Source-Dependent Modulation of Leukocyte Adhesion to Primary Microvascular Endothelial Cells

Anti-inflammatory effects of low-dose irradiation often follow a non-linear dose–effect relationship. These characteristics were also described for the modulation of leukocyte adhesion to endothelial cells. Previous results further revealed a contribution of reactive oxygen species (ROS) and anti-oxidative factors to a reduced leukocyte adhesion. Here, we evaluated the expression of anti-oxidative enzymes and the transcription factor Nrf2 (Nuclear factor-erythroid-2-related factor 2), intracellular ROS content, and leukocyte adhesion in primary human microvascular endothelial cells (HMVEC) upon low-dose irradiation under physiological laminar shear stress or static conditions after irradiation with X-ray or Carbon (C)-ions (0–2 Gy). Laminar conditions contributed to increased mRNA expression of anti-oxidative factors and reduced ROS in HMVEC following a 0.1 Gy X-ray and 0.5 Gy C-ion exposure, corresponding to reduced leukocyte adhesion and expression of adhesion molecules. By contrast, mRNA expression of anti-oxidative markers and adhesion molecules, ROS, and leukocyte adhesion were not altered by irradiation under static conditions. In conclusion, irradiation of endothelial cells with low doses under physiological laminar conditions modulates the mRNA expression of key factors of the anti-oxidative system, the intracellular ROS contents of which contribute at least in part to leucocyte adhesion, dependent on the radiation source.

Improved cellular automata model shows that indirect apoptotic cell death due to vascular damage enhances the local control of tumors by single fraction high-dose irradiation

We investigated the effects of indirect apoptotic cell death due to vascular damage on tumor response to a single large dose with an improved two-dimensional cellular automata model. The tumor growth was simulated by considering the oxygen and nutrients supplied to the tumor through the blood vessels. The cell damage processes were modeled by taking account of the direct cell death and the indirect death due to the radiation-induced vascular damages. The radiation increased the permeation of oxygen and nutrients through the blood vessel or caused the breakdown of the vasculature. The amount of oxygen in cancer cells affected the response of cancer cells to radiation and the tumor growth rate after irradiation. The lack of oxygen led to the apoptotic death of cancer cells. We calculated the tumor control probability (TCP) at different radiation doses, D, the probability of apoptotic death, PO2_ap, the threshold of the oxygen level for indirect apoptotic death, O2t, the average oxygen level in cancer cells, [O2]av, and the vessel survival probability after radiation damage, Pv. Due to the vessel damage, indirect cell death led to a 4% increase in TCP for the dose ranging from 15 Gy to 20 Gy. TCP increased with increasing PO2_ap and O2t due to increased apoptotic death. The variation of TCP as a function of [O2]av exhibited the minimum at [O2]av of 2.7%. The apoptosis increased as [O2]av decreased, leading to an increasing TCP. On the other hand, the direct radiation damage increased, and the apoptosis decreased for higher [O2]av, resulting in a higher TCP. We showed by modeling the radiation damage of blood vessels in a 2D CA simulation that the indirect apoptotic death of cancer cells, caused by the reduction of the oxygen level due to vascular damage after high dose irradiation, increased TCP.

MPC-6 Clinical significance of whole chromosomal aberration signatures in non-metastatic medulloblastomas treated with 18Gy of craniospinal irradiation

Background: One of the most significant challenges is a reduction in the dose of craniospinal irradiation (CSI) in patients with medulloblastoma to minimize neurological sequelae. However, a North American clinical trial failed to show the prognostic non-inferiority of lower-dose irradiation compared to that associated with standard-dose radiation therapy for non-metastatic medulloblastomas. A European retrospective study revealed that whole chromosomal aberration signatures (WCASs) are a potential prognostic factor in Group 3/4 medulloblastoma without metastasis, but whether the molecular signature has the same clinical impact in patients treated with lower-dose CSI remains unknown. Methods: We conducted DNA methylation analysis using an Illumina Infinium Human Methylation EPIC BeadChip array to investigate molecular prognostic markers in 23 medulloblastoma patients who were registered in the Japan Pediatric Molecular Neuro-Oncology Group and treated with lower-dose CSI relative to standard treatment. A WCAS was defined as the presence of at least two of three chromosomal changes as follows: chromosome (chr) 7 gain, chr 8 loss, and chr 11 gain.Results: All patients presented with no residue or a residual tumor smaller than 1.5 cm2 after surgery without metastasis. The median age at onset was 6.9 years, and the median follow-up period was 80.6 months. CSI was delivered at a median dose of 18.0 Gy. Regarding molecular subgrouping, there were 5 WNT, 2 SHH, 1 Group 3, and 15 Group 4 medulloblastomas. Seven patients with Group 3/4 medulloblastomas showed WCASs and had significantly better prognosis than those without the alteration (5-year progression-free survival 100% vs. 63%, p = 0.046). Two late relapses occurred at 89 and 115 months after diagnosis, respectively, and one of these patients presented with a WCAS.Conclusion: WCAS may be a molecular prognostic marker not only in patients with medulloblastoma treated with standard-dose CSI but also in those treated with lower-dose irradiation.

Assessment of the photosensitizer efficacy after determination of threshold doses initiating erythrocyte hemolysis

Purpose: To find out a technique for determining the effectiveness of porphyrin-type photosensitizers with concentrations which cause hemolysis in erythrocytes (hemolysis threshold). The hemolysis threshold is found under the following conditions: irradiation in the Soret band with standard parameters – power density, exposure dose, irradiation time.Material and methods. Phototherapeutic device “AST” (LLC “Pankov-medical”) – average power 0.5 W, wavelength ≈ 405 nm. Rat blood erythrocytes. “Multiscan MS” device manufactured by Labsystems, Finland. Preparation “Dimegin”, hematoporphyrin derivative; preparation “Photoditazine” (LLC “VETA-GRAND”), chlorine E6 derivative.Results. On analyzing the results obtained after experimentation with the abovementioned two preparations, it was shown that the developed technique has a high efficacy. It was also shown that the studied photosensitizers are equally effective when irradiated in the Soret band.Conclusion. The developed technique is simple and easy to use. It helps to assess the efficacy of various photosensitizers thus, facilitating the selection of their necessary concentrations for managing different diseases.

Low dose irradiation of plum peel anthocyanins: Impact on stability and antioxidant activity

Consumer demand for both natural food colorants and non-thermal methods of preservation has increased overwhelmingly during the last few decades. Preventing loss of appeal and color through non-thermal methods without compromising on quality will provide an alternative to conventional thermal methods. In the present study, irradiation of plum peel anthocyanins after extraction and purification was done in a range of 0.1-1.0 kGy. Results revealed that degradation was significantly (p ≤ 0.05) dependent on both time and dose of irradiation. The percentage degradation observed in the first 3 h of treatment was 9.0-21.0±1.2%. The maximum anthocyanin degradation recorded during 8 h of irradiation was 45.3%. Anthocyanin degradation was apparently less at 0.1-0.5 kGy compared to 0.6-1.0 kGy. The retention of anthocyanins was up to the extent of 88.8-62.3% in samples irradiated at 0.1-0.5 kGy. Comparison of the individual anthocyanin indicated that diglucoside anthocyanins were stable towards irradiation than monoglucoside anthocyanins. For monoglucoside anthocyanins, a 50% decrease was observed as the irradiation treatment exceeded 0.6 kGy. A close comparison of the data revealed that at irradiation dose of 1.0 kGy, the decrease in L* and a* values was almost four and three times higher than that observed in samples irradiated at 0.5 kGy and below; while an increase in b* value was almost double. In spite of marginal degradation in color, low dose irradiation (up to 0.5 kGy) significantly (p≤0.05) increased the antioxidant activity of the plum peel anthocyanins.