Elette Engels

Purpose DNA incorporated bromodeoxyuridine (BrUdR) can locally enhance tumor damage when irradiated with the optimal energy for the Z of bromine in BrUdR. This research investigates the potential for improved treatment outcomes for brain cancer patients by combining the current standard treatment of radiotherapy and Temozolomide (TMZ) with DNA incorporated BrUdR and irradiation with megavoltage, kilovoltage or synchrotron radiations.Materials and methods Clonogenic assay was the endpoint for the assessment of 9LGS gliosarcoma cells pre-exposed to TMZ or BrUdR singly or co-administered. 9LGS cells were irradiated with x-rays at either clinically available low dose rate megavoltage, or orthovoltage and ultra-high dose rate synchrotron radiations. Results were supported by cell cycle, BrUdR uptake, senescence, and γH2AX double strand break analysis.Results BrUdR incorporation increased when combined with TMZ compared to 9LGS treated with BrUdR alone. For all beams tested, increased cell death was observed for 9LGS treated with BrUdR and BrUdR + TMZ with greatest radiosensitivity for 9LGS pre-exposed to BrUdR + TMZ. This correlated with additional incorporation of BrUdR and more cells in the radiosensitive G2/M phase. Maximum radiosensitivity was observed for 9LGS treated with BrUdR + TMZ and irradiated at an ultra-high dose rate.Conclusion This study demonstrated significant and synergistic, increases in cell death are achievable when BrUdR + TMZ treated cells are irradiated with ultra-high dose rate synchrotron radiation, highlighting the potential of this novel combination to treat brain cancer. Of equal significance is that co-administration of BrUdR and TMZ increased cell death with 6 MV irradiation.

Objective: This study aims to develop canine radiotherapy phantoms and a dosimetry protocol for synchrotron microbeam radiation therapy (MRT). Approach: Two canine head phantoms were designed based on computed tomography (CT) image sets obtained from veterinary patients. Polylactic acid polymer and calcium sulfate hemihydrate plaster were used to radiologically simulate soft tissue and bone. Fused deposition modeling 3D printing was used to fabricate the external geometry of the head phantoms. CT images of the phantoms were acquired to measure its CT number profile and to create treatment plans. Using a 4 Tesla superconducting multipole wiggler field and in vacuo filters, synchrotron beam was delivered as broad-beam and as microbeam arrays at the imaging and medical beamline of the Australian Synchrotron. The Monte Carlo simulated dose using a hybrid EclipseTM treatment planning system (TPS) was compared to the experimental measurements. Main results: The head phantoms offer a realistic three-dimensional canine head geometry and CT numbers mimicing soft tissue and bone. Various types of detectors and biological samples can be accomodated, using a series of customizable rectangular inserts. The measured accumulated doses for microbeam irradiation were within +/- 2% of the TPS doses, while all the other measured doses were within +/- 10% agreement with the TPS doses. The TPS, phantom positioning, and detector uncertainties contributed to the measurement discrepancies. Significance and conclusions: The 3D printed canine phantoms were shown to be useful dosimetry tool for synchrotron MRT end-to-end quality assurance. The development of canine phantoms and dosimetry methodology are critical for the treatment plan optimization required in future veterinary studies. Both the phantoms and the dosimetry protocol are valuable tools in future veterinary MRT trials.

Very-high-energy electrons, coupled with ultra-high dose rates, are being explored for their potential use in radiotherapy to treat deep-seated tumours. The dose per pulse needed to achieve ultra-high dose rates far exceeds the limit of current medical linear accelerator capabilities. A high dose per pulse has been observed as the limiting factor for many existing dosimeters, resulting in saturation at doses far below what is required. The MOSkin, an existing clinical quality assurance dosimeter, has previously been demonstrated as dose rate independent but has not been subjected to a high dose per pulse. Within this study, the MOSkins dose-per-pulse response was tested for linearity, with a dose per pulse as high as 23 Gy within 200 ns at the ANSTO Australian Synchrotron’s Pulsed Energetic Electrons for Research facility. While using EBT-XD film as a reference dosimeter, a dose rate dependence of the EBT-XD was discovered. Once confirmed and a correction factor established, EBT-XD was used as an independent reference measurement. This work presents confirmation of the MOSkin suitability for ultra-high dose-rate environments with an electron energy of 100 MeV, and a theoretical discussion of its dose-rate and dose-per-pulse independence; the MOSkin is the only detector suitable for both clinical quality assurance, and ultra-high dose-rate measurements in its standard, unmodified form.

The results of radiotherapy in patients with primary malignant brain tumors are extremely dissatisfactory: the overall survival after a diagnosis of glioblastoma is typically less than three years. The development of spatially fractionated radiotherapy techniques could help to improve this bleak prognosis. In order to develop technical equipment and organ-specific therapy plans, dosimetry studies as well as radiobiology studies are conducted. Although perfect spheres are considered optimal phantoms by physicists, this does not reflect the wide variety of head sizes and shapes in our patient community. Depth from surface and X-ray dose absorption by tissue between dose entry point and target, two key parameters in medical physics planning, are largely determined by the shape and thickness of the skull bone. We have, therefore, designed and produced a biomimetic tool to correlate measured technical dose and biological response in human cancer cells: a brain phantom, produced from tissue-equivalent materials. In a first pilot study, utilizing our phantom to correlate technical dose measurements and metabolic response to radiation in human cancer cell lines, we demonstrate why an anthropomorphic phantom is preferable over a simple spheroid phantom.

High-Z nanoparticles (NPs) have the potential to revolutionize cancer radiotherapy by radiosensitising tumours. This is particularly important for radioresistant cancers such as glioblastoma. A newer NP candidate in this area is thulium oxide nanoparticles (TmNPs). However, prior to clinical assessment, ideal NP characteristics, including biocompatibility, biosafety, and preferential uptake in cancer, should be assessed. This in vitro study compares the effects of TmNP treatment, without radiation, on 9L gliosarcoma (9LGS), a well-established glioblastoma cell model, with exposure to Madin Darby Canine Kidney (MDCK) cells, a widely used non-cancerous cell model. The findings demonstrated selective uptake of TmNPs in 9LGS over MDCK following treatment. A biological assessment of toxicity confirmed minimal long-term effects on MDCK, whilst TmNPs were observed to induce some notable cell death in 9LGS. Excessive TmNP uptake in 9LGS over time was observed to induce cell vacuolisation, which resulted in cell death via necrosis. It was concluded that this was the explanation for the underlying mechanisms of TmNP toxicity in cancer cells. This study was therefore able to demonstrate not only that TmNPs are a biocompatible, cancer-selective candidate for radiosensitiser usage, but further provided a theory to explain its mechanisms of cancer cell toxicity.

Background/objectives: Brain cancer remains difficult to treat, with survival statistics stagnant for decades. The resistance of glioblastoma brain tumours can greatly challenge the effectiveness of conventional cancer radiotherapy. However, high dose rate radiotherapy has unique effects that allow for normal tissue sparing whilst maintaining tumour control. The addition of targeted radiosensitisers, such as the chemotherapeutic drug methotrexate (MTX) or the high-Z halogenated pyrimidine drug iododeoxyuridine (IUdR), can improve radiotherapy outcomes. Combining these radiosensitiser agents with ultra-high dose rate (UHDR) synchrotron X-rays can bear synergistic effects to enhance the efficacy of these multi-modal UHDR therapies, providing a means to overcome the radioresistance of brain cancer. Methods: Here, we use controlled in vitro assays following treatment, including a clonogenic assay to determine long-term cell survival and γH2AX immunofluorescent confocal microscopy to quantify double-strand DNA breaks (DSBs). Results: We find significant enhancement for highly synergistic combinations of IUdR+MTX with synchrotron X-rays. Cell survival results demonstrate 5.4 times increased 9L gliosarcoma cell killing when these agents are combined with UHDR synchrotron X-rays compared with conventional X-rays alone at the same 5 Gy dose. The underlying mechanisms are unveiled using γH2AX imaging and reveal significant increases in DSBs and dying cells following exposure to UHDR radiation. Conclusions: Our results demonstrate that highly synergistic combination treatments using UHDR synchrotron radiation can yield significantly improved brain cancer killing compared with conventional radiotherapy. We anticipate that these additive, multi-modal combination therapies will provide options for more targeted and effective use of radiotherapies for the future treatment of brain cancer.

Background/Objectives: Brain cancer is notoriously resistant to traditional treatments, including radiotherapy. Microbeam radiation therapy (MRT), arrays of ultra-fast synchrotron X-ray beams tens of micrometres wide (called peaks) and spaced hundreds of micrometres apart (valleys), is an effective alternative to conventional treatments. MRT’s advantage is that normal tissues can be spared from harm whilst maintaining tumour control. Combining MRT with targeted radiosensitisers, such as nanoparticles, chemotherapeutic drugs, and halogenated pyrimidine drugs, can further improve radiotherapy by enhancing radiation damage. However, the underlying mechanisms of MRT are still being understood, which is essential to ensuring the reliable and successful use of MRT. Methods: An in vitro study was performed using γH2AX imaging, and quantification was performed via confocal microscopy and a clonogenic cell survival assay. Results: We show that methotrexate chemotherapeutics and iododeoxyuridine enhance MRT cell-killing and thulium oxide nanoparticles (TmNPs) broaden MRT peaks, and using γH2AX immunofluorescent confocal microscopy to quantify DNA damage, we further our knowledge of MRT mechanisms. γH2AX images verify the biological responses of cells aligning with the physical collimation of MRT, and we can accurately measure MRT microbeam characteristics bio-dosimetrically. The peak-to-valley dose ratio (PVDR), the ratio of the peak dose to the valley dose that characterises an MRT field, was accurately measured biologically using γH2AX imaging, despite studies previously finding this challenging. Conclusions: The measurement of biological PVDR has been performed for the first time with high-Z radiosensitisers, including nanoparticles, and several novel radiosensitiser-enhanced MRT mechanisms were discovered. Our results deepen our understanding of MRT with radiosensitisers, and can contribute to its accurate and future successful use in treating cancer.

Objective. This study aims to design, manufacture, and test 3D printed quality assurance (QA) dosimetry phantoms for synchrotron brain cancer radiation therapy at the Australian synchrotron. Approach. Fabricated 3D printed phantoms from simple slab phantoms, a preclinical rat phantom, and an anthropomorphic head phantom were fabricated and characterized. Attenuation measurements of various polymers, ceramics and metals were acquired using synchrotron monochromatic micro-computed tomography (CT) imaging. Polylactic acid plus, VeroClear, Durable resin, and tricalcium phosphate were used in constructing the phantoms. Furthermore, 3D printed bone equivalent materials were compared relative to ICRU bone and hemihydrate plaster. Homogeneous and heterogeneous rat phantoms were designed and fabricated using tissue-equivalent materials. Geometric accuracy, CT imaging, and consistency were considered. Moreover, synchrotron broad-beam x-rays were delivered using a 3 Tesla superconducting multipole wiggler field for four sets of synchrotron radiation beam qualities. Dose measurements were acquired using a PinPoint ionization chamber and compared relative to a water phantom and a RMI457 Solid Water phantom. Experimental depth doses were compared relative to calculated doses using a Geant4 Monte Carlo simulation. Main results. Polylactic acid (PLA+) shows to have a good match with the attenuation coefficient of ICRU water, while both tricalcium phosphate and hydroxyapatite have good attenuation similarity with ICRU bone cortical. PLA+ material can be used as substitute to RMI457 slabs for reference dosimetry with a maximum difference of 1.84%. Percent depth dose measurement also shows that PLA+ has the best match with water and RMI457 within ±2.2% and ±1.6%, respectively. Overall, PLA+ phantoms match with RMI457 phantoms within ±3%. Significance and conclusion. The fabricated phantoms are excellent tissue equivalent equipment for synchrotron radiation dosimetry QA measurement. Both the rat and the anthropomorphic head phantoms are useful in synchrotron brain cancer radiotherapy dosimetry, experiments, and future clinical translation of synchrotron radiotherapy and imaging.