Large Field Alpha Irradiation Setup for Radiobiological Experiments

Roobol, Stefan J.; Kouwenberg, J.J.M.; Denkova, Antonia G.; Kanaar, Roland; Essers, Jeroen

doi:10.3390/mps2030075

Cited by 6 publications

(3 citation statements)

References 29 publications

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“…X-ray irradiation was performed using the RS320 (Xstrahl Live Sciences), a self-contained cabinet, with a dose rate of 1.6554 Gy/min and working voltage of 195 kV and 10 mA. Alpha irradiation was performed as described before [46]. In brief, coverslips containing cultured cells were washed with PBS and placed upside down (cells facing down) on a Mylar dish.…”

Section: X-ray and α-Particle Irradiationmentioning

confidence: 99%

Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells

Roobol

Bent

Cappellen

et al. 2020

IJMS

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High-linear-energy-transfer (LET) radiation is more lethal than similar doses of low-LET radiation types, probably a result of the condensed energy deposition pattern of high-LET radiation. Here, we compare high-LET α-particle to low-LET X-ray irradiation and monitor double-strand break (DSB) processing. Live-cell microscopy was used to monitor DNA double-strand breaks (DSBs), marked by p53-binding protein 1 (53BP1). In addition, the accumulation of the endogenous 53BP1 and replication protein A (RPA) DSB processing proteins was analyzed by immunofluorescence. In contrast to α-particle-induced 53BP1 foci, X-ray-induced foci were resolved quickly and more dynamically as they showed an increase in 53BP1 protein accumulation and size. In addition, the number of individual 53BP1 and RPA foci was higher after X-ray irradiation, while focus intensity was higher after α-particle irradiation. Interestingly, 53BP1 foci induced by α-particles contained multiple RPA foci, suggesting multiple individual resection events, which was not observed after X-ray irradiation. We conclude that high-LET α-particles cause closely interspaced DSBs leading to high local concentrations of repair proteins. Our results point toward a change in DNA damage processing toward DNA end-resection and homologous recombination, possibly due to the depletion of soluble protein in the nucleoplasm. The combination of closely interspaced DSBs and perturbed DNA damage processing could be an explanation for the increased relative biological effectiveness (RBE) of high-LET α-particles compared to X-ray irradiation.

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Section: X-ray and α-Particle Irradiationmentioning

confidence: 99%

Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells

Roobol

Bent

Cappellen

et al. 2020

IJMS

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“…Similar observations have been made in cells irradiated with other particles, such as α-particles and carbon ions ( Nagle et al, 2016 ; Roobol et al, 2020 ). 53BP1 foci induced by α-particle are bigger than foci induced by photons ( Roobol et al, 2019 ). Following live dynamics of GFP-tagged 53BP1 foci in α-particle irradiated cells has shown that the repair of DSBs is slower after high LET IR compared to low LET IR.…”

Section: Induction Of Dna Damage By Different Types Of Radiationmentioning

confidence: 98%

DNA Double Strand Break Repair Pathways in Response to Different Types of Ionizing Radiation

et al. 2021

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The superior dose distribution of particle radiation compared to photon radiation makes it a promising therapy for the treatment of tumors. However, the cellular responses to particle therapy and especially the DNA damage response (DDR) is not well characterized. Compared to photons, particles are thought to induce more closely spaced DNA lesions instead of isolated lesions. How this different spatial configuration of the DNA damage directs DNA repair pathway usage, is subject of current investigations. In this review, we describe recent insights into induction of DNA damage by particle radiation and how this shapes DNA end processing and subsequent DNA repair mechanisms. Additionally, we give an overview of promising DDR targets to improve particle therapy.

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“…Plasmid and construct information is available on request. Transfected cells were externally irradiated using previously described methods [34]. Live cell imaging was performed on a confocal microscope (Leica SP5) overnight (16 hours).…”

Section: Confocal (Live Cell) Microscopymentioning

confidence: 99%

Uptake and subcellular distribution of radiolabeled polymersomes for radiotherapy

Roobol¹,

Hartjes²,

Slotman³

et al. 2020

Nanotheranostics

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Polymersomes have the potential to be applied in targeted alpha radionuclide therapy, while in addition preventing release of recoiling daughter isotopes. In this study, we investigated the cellular uptake, post uptake processing and intracellular localization of polymersomes. Methods: High-content microscopy was used to validate polymersome uptake kinetics. Confocal (live cell) microscopy was used to elucidate the uptake mechanism and DNA damage induction. Intracellular distribution of polymersomes in 3-D was determined using super-resolution microscopy. Results: We found that altering polymersome size and concentration affects the initial uptake and overall uptake capacity; uptake efficiency and eventual plateau levels varied between cell lines; and mitotic cells show increased uptake. Intracellular polymersomes were transported along microtubules in a fast and dynamic manner. Endocytic uptake of polymersomes was evidenced through co-localization with endocytic pathway components. Finally, we show the intracellular distribution of polymersomes in 3-D and DNA damage inducing capabilities of 213 Bi labeled polymersomes. Conclusion: Polymersome size and concentration affect the uptake efficiency, which also varies for different cell types. In addition, we present advanced assays to investigate uptake characteristics in detail, a necessity for optimization of nano-carriers. Moreover, by elucidating the uptake mechanism, as well as uptake extent and geometrical distribution of radiolabeled polymersomes we provide insight on how to improve polymersome design.

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Large Field Alpha Irradiation Setup for Radiobiological Experiments

Cited by 6 publications

References 29 publications

Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells

Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells

DNA Double Strand Break Repair Pathways in Response to Different Types of Ionizing Radiation

Uptake and subcellular distribution of radiolabeled polymersomes for radiotherapy

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