Commissioning of a RayStation structure template for the iBEAM evo Couchtop.

Accurate radiotherapy treatment planning requires attenuation through the treatment couch to be accounted for in dose calculation. This is commonly performed by using contouring tools to add a virtual structure in the shape of the treatment couch and assigning the preferred absorption properties. The RayStation treatment planning system (TPS) allows users to assign a material that comprises both an elemental structure and a physical density. The selection of such parameters should be made so that modelled attenuation through the couch closely matches measured data. When these measurements involve the use of plastic phantoms and rotational beams, the validity of the data is dependent upon aspects of TPS and linear accelerator performance that can be difficult to quantify. A fundamental measure of couch attenuation using an ionisation chamber in water and perpendicular beam geometry that required no gantry movement was implemented to eliminate the identified uncertainties. This data was used to determine the combination of elemental composition and density assigned to a modelled couch structure that provided the most accurate representation of beam attenuation in this simple geometry. The preferred material was then validated using a cylindrical phantom and rotational beams. The findings were equivalent between the static gantry with water phantom and rotating gantry with cylindrical phantom. Of the elemental compositions investigated, it was possible to achieve suitable agreement with the measured data for each option provided the density was optimised. Choice of the elemental composition was not observed to be an important factor in achieving a good model.

Targeting of externalized αB-crystallin on irradiated endothelial cells with pro-thrombotic vascular targeting agents: Potential applications for brain arteriovenous malformations.

BACKGROUND: Vascular targeting uses molecular markers on the surface of diseased vasculature for ligand-directed drug delivery to induce vessel occlusion or destruction. In the absence of discriminatory markers, such as in brain arteriovenous malformations (AVMs), stereotactic radiosurgery may be used to prime molecular changes on the endothelial surface. This study explored αB-crystallin (CRYAB) as a radiation induced target and pre-tested the specificity and efficacy of a CRYAB-targeting coaguligand for in vitro thrombus induction. METHODS: A parallel-plate flow system was established to circulate human whole blood over a layer of human brain endothelial cells. A conjugate of anti-CRYAB antibody and thrombin was injected into the circuit to compare binding and thrombus formation on cells with or without prior radiation treatment (0-25 Gy). RESULTS: Radiation increased CRYAB expression and surface exposure in human brain endothelial cells. In the parallel-plate flow system, the targeted anti-CRYAB-thrombin conjugate increased thrombus formation on the surface of irradiated cells relative to non-irradiated cells and to a non-targeting IgG-thrombin conjugate. Fibrin deposition and accumulation of fibrinogen degradation products increased significantly at radiation doses at or above 15 Gy with conjugate concentrations of 1.25 and 2.5 µg/mL. CONCLUSIONS: CRYAB exposure can be detected at the surface of human brain endothelial cells in response to irradiation. Pro-thrombotic CRYAB-targeting conjugates can bind under high flow conditions and in the presence of whole blood induce stable thrombus formation with high specificity and efficacy on irradiated surfaces. CRYAB provides a novel radiation marker for potential vascular targeting in irradiated brain AVMs.

Occlusion of Animal Model Arteriovenous Malformations Using Vascular Targeting.

Brain arteriovenous malformations (AVMs) are a significant cause of intracerebral hemorrhage in children and young adults. Currently, one third of patients have no viable treatment options. Vascular targeting agents (VTAs) are being designed to deliver pro-thrombotic molecules to the abnormal AVM vessels for rapid occlusion and cure. This study assessed the efficacy of a pro-thrombotic VTA targeting phosphatidylserine (PS) in a radiation-primed AVM animal model. The model AVM was surgically created in rats by anastomosis of the left external jugular vein to the adjacent common carotid artery. After 6 weeks, the AVM was irradiated (20 Gy) using gamma knife surgery (GKS). A PS-targeting VTA was created by conjugation of annexin V with human thrombin and administered intravenously 3 weeks post-GKS or sham. Unconjugated thrombin was used as a non-targeting control. AVM thrombosis and occlusion was monitored 3 weeks later by angiography and histology. Preliminary experiments established a safe dose of active thrombin for systemic administration. Subsequently, a single dose of annexin V-thrombin conjugate (0.77 mg/kg) resulted in angiographic AVM occlusion in sham (75%) and irradiated (63%) animals, while non-targeted thrombin did not. Lowering the conjugate dose (0.38 mg/kg) decreased angiographic AVM occlusion in sham (13%) relative to irradiated (80%) animals (p = 0.03) as did delivery of two consecutive doses of 0.38 mg/kg, 2 days apart (sham (0%); irradiated (78%); p = 0.003). These findings demonstrate efficacy of the PS-targeting VTA and the feasibility of a vascular targeting approach for occlusion of high-flow AVMs. Targeting specificity can be enhanced by radiation-sensitization and VTA dose modification.

Radiation-Stimulated Translocation of CD166 and CRYAB to the Endothelial Surface Provides Potential Vascular Targets on Irradiated Brain Arteriovenous Malformations.

Vascular targeting with pro-thrombotic antibody-conjugates is a promising biological treatment for brain arteriovenous malformations (bAVMs). However, targeted drug delivery relies on the identification of unique or overexpressed markers on the surface of a target cell. In the absence of inherent biological markers, stereotactic radiosurgery may be used to prime induction of site-specific and targetable molecular changes on the endothelial surface. To investigate lumen-accessible, endothelial targets induced by radiation, we combined Gamma knife surgery in an AVM animal model with in vivo biotin-labeling and comparative proteomics. Two proteins, αB-crystallin (CRYAB)-a small heat shock protein that normally acts as an intracellular chaperone to misfolded proteins-and activated leukocyte cell adhesion molecule CD166, were further validated for endothelial surface expression after irradiation. Immunostaining of endothelial cells in vitro and rat AVM tissue ex vivo confirmed de novo induction of CRYAB following irradiation (20 Gy). Western analysis demonstrated that CRYAB accumulated intracellularly as a 20 kDa monomer, but, at the cell surface, a novel 65 kDa protein was observed, suggesting radiation stimulates translocation of an atypical CRYAB isoform. In contrast, CD166 had relatively high expression in non-irradiated cells, localized predominantly to the lateral surfaces. Radiation increased CD166 surface exposure by inducing translocation from intercellular junctions to the apical surface without significantly altering total protein levels. These findings reinforce the dynamic molecular changes induced by radiation exposure, particularly at the cell surface, and support further investigation of radiation as a priming mechanism and these molecules as putative targets for focused drug delivery in irradiated tissue.

Volume not number of metastases: Gamma Knife radiosurgery management of intracranial lesions from an Australian perspective.

BACKGROUND AND PURPOSE: To assess the response of the first cohort of patients treated with Gamma Knife radiosurgery in Australia. MATERIALS AND METHODS: A prospectively collected cohort of 180 patients with intracranial metastases from different primaries was treated between August 2010 and July 2017. Survival was calculated using the Kaplan-Meier's method. Cox regression was used for multivariate analysis. RESULTS: Currently 141 patients (78.3%) have died of their disease. The median survival for the group as a whole was 9.2 months, with observed differences resulting from the volume of tumor burden (11.4 months for volumes <3.2 cm3 to 5.16 months for volume >9.1 cm3). Overall 2-year survival was 20.7%. CONCLUSION: Results from the first Gamma Knife radiosurgery center in Australia showed that the treatment is feasible and effective, consistent with the international experience. For patients with larger numbers of intracranial metastases, the total volume of the intracranial burden may be of more significance in predicting outcomes. While there appeared to be a difference in survival by histologic origin, this could be related to concurrent systemic immunotherapy available for certain tumors.

The Elekta Fraxion™ system is not suitable for maxillary fixation in canine conformal radiation therapy techniques.

In this prospective, exploratory study, we evaluated the positioning accuracy in a group of 15 dogs undergoing fractionated stereotactic radiotherapy for tumors affecting the head, using a modified human maxillary fixation device (Elekta Fraxion™ system). Positioning was assessed using on-board volumetric imaging, with a six-degrees-of-freedom image registration technique. Prior to treatment delivery, CBCT images were obtained and patient alignment was corrected, in both translational and rotational planes, using a six-degrees-of-freedom robotic patient positioning system (HexaPOD Evo RT System). The maximum angular inter-fraction motions observed were 6.1° (yaw), 10.9° (pitch), and 4.5° (roll). The mean systematic translational errors were 4.7, 2.6, and 2.3 mm, mean random translational errors were 3.0, 2.2, and 2.5 mm, and mean overall translational errors were 2.4, 0.7, and 2.3 mm in the cranial-caudal, lateral, and dorsal-ventral directions, respectively. The mean systematic rotational errors were 1.17°, 0.77°, and 1.43°, the mean rotational random errors were 1.65°, 1.46°, and 1.34° and the mean overall rotational errors were 0.56°, 0.22°, and 0.29° in the yaw, pitch, and roll directions, respectively. The mean error of the three-dimensional vector was 6.9 mm with a standard deviation of 3.8 mm. Ninety-five percent of the three-dimensional vectors were <14.8 mm. This study demonstrates that this maxillary fixation device relies on six-degrees-of-freedom registration and an ability to apply corrections using a six-degrees-of-freedom couch for accurate patient positioning and tumor targeting. Its use in conformal radiation therapy in dogs is not recommended.

Effects of FOXM1 inhibition and ionizing radiation on melanoma cells.

Metastatic melanoma can be highly refractory to conventional radiotherapy and chemotherapy but combinatorial-targeted therapeutics are showing greater promise on improving treatment efficacy. Previous studies have shown that knockdown of Forkhead box M1 (FOXM1) can sensitize various tumor types to radiation-induced cell death. The effect of combining radiation with a small molecule FOXM1 inhibitor, Siomycin A, on growth, death and migration of a metastatic melanoma cell line (SK-MEL-28) that overexpresses this pleiotropic cell cycle regulator was investigated. Siomycin A (SIOA) was found to be a strong inducer of apoptosis, and inhibitor of proliferation and migration in a scratch wound assay in this cell line. Induction of apoptosis occurred at concentrations >1 µM in association with reductions in the constitutive FOXM1 and anti-apoptotic B-cell lymphoma 2 protein levels found in these cells. Single doses of ionizing radiation (0-40 Gy) delivered by linear accelerator caused inhibition of growth and migration without significant induction of cell death. Pretreatment with SIOA did not increase the sensitivity of this melanoma cell line to radiation as observed in other tumor types. These data confirm that as a single agent, SIOA is an effective inducer of cell death and inhibitor of migration in metastatic melanoma cells expressing constitutive FOXM1. In combination with radiation, SIOA pre-treatment, however, may not be of added benefit.

Ionizing radiation reduces ADAM10 expression in brain microvascular endothelial cells undergoing stress-induced senescence.

Cellular senescence is associated with aging and is considered a potential contributor to age-associated neurodegenerative disease. Exposure to ionizing radiation increases the risk of developing premature neurovascular degeneration and dementia but also induces premature senescence. As cells of the cerebrovascular endothelium are particularly susceptible to radiation and play an important role in brain homeostasis, we investigated radiation-induced senescence in brain microvascular endothelial cells (EC). Using biotinylation to label surface proteins, streptavidin enrichment and proteomic analysis, we analyzed the surface proteome of stress-induced senescent EC in culture. An array of both recognized and novel senescence-associated proteins were identified. Most notably, we identified and validated the novel radiation-stimulated down-regulation of the protease, a disintegrin and metalloprotease 10 (ADAM10). ADAM10 is an important modulator of amyloid beta protein production, accumulation of which is central to the pathologies of Alzheimer's disease and cerebral amyloid angiopathy. Concurrently, we identified and validated increased surface expression of ADAM10 proteolytic targets with roles in neural proliferation and survival, inflammation and immune activation (L1CAM, NEO1, NEST, TLR2, DDX58). ADAM10 may be a key molecule linking radiation, senescence and endothelial dysfunction with increased risk of premature neurodegenerative diseases normally associated with aging.

Radiosurgery Alters the Endothelial Surface Proteome: Externalized Intracellular Molecules as Potential Vascular Targets in Irradiated Brain Arteriovenous Malformations.

Stereotactic radiosurgery (SRS) is an established treatment for brain arteriovenous malformations (AVMs) that drives blood vessel closure through cellular proliferation, thrombosis and fibrosis, but is limited by a delay to occlusion of 2-3 years and a maximum treatable size of 3 cm. In this current study we used SRS as a priming tool to elicit novel protein expression on the endothelium of irradiated AVM vessels, and these proteins were then targeted with prothrombotic conjugates to induce rapid thrombosis and vessel closure. SRS-induced protein changes on the endothelium in an animal model of AVM were examined using in vivo biotin labeling of surface-accessible proteins and comparative proteomics. LC-MS/MS using SWATH acquisition label-free mass spectrometry identified 280 proteins in biotin-enriched fractions. The abundance of 56 proteins increased after irradiation of the rat arteriovenous fistula (20 Gy, ≥1.5-fold). A large proportion of intracellular proteins were present in this subset: 29 mitochondrial and 9 cytoskeletal. Three of these proteins were chosen for further validation based on previously published evidence for surface localization and a role in autoimmune stimulation: cardiac troponin I (TNNI3); manganese superoxide dismutase (SOD2); and the E2 subunit of the pyruvate dehydrogenase complex (PDCE2). Immunostaining of AVM vessels confirmed an increase in abundance of PDCE2 across the vessel wall, but not a measurable increase in TNNI3 or SOD2. All three proteins co-localized with the endothelium after irradiation, however, more detailed subcellular distribution could not be accurately established. In vitro, radiation-stimulated surface translocation of all three proteins was confirmed in nonpermeabilized brain endothelial cells using immunocytochemistry. Total protein abundance increased modestly after irradiation for PDCE2 and SOD2 but decreased for TNNI3, suggesting that radiation primarily affects subcellular distribution rather than protein levels. The novel identification of these proteins as surface exposed in response to radiation raises important questions about their potential role in radiation-induced inflammation, fibrosis and autoimmunity, but may also provide unique candidates for vascular targeting in brain AVMs and other vascular tissues.

Evaluation of accuracy and reproducibility of a relocatable maxillary fixation system for fractionated intracranial stereotactic radiation therapy.

INTRODUCTION: Accurate localisation is an essential component for the delivery of intracranial stereotactic treatment. For fractionated stereotactic radiotherapy, we compared the daily localisation accuracy of a standard thermoplastic mask with a new maxillary fixation device (MFD). METHODS: Daily pre-treatment kV cone-beam computed tomography (CBCT) scans of 23 patients (12 localised in the MFD and 11 in the mask) with benign skull-based lesions were reviewed retrospectively. The set up accuracy was measured in 6° of freedom, to ascertain both individual and population random and systematic errors. The appropriate clinical target volume to planning target volume margin was computed from set up error data. RESULTS: A total of 682 CBCT scans were evaluated. Systematic (Σ) and random (σ) population errors were Σ = 0.8 mm, 0.2 mm and 0.2 mm and σ = 0.3 mm, 0.3 mm and 0.2 mm, respectively, for the standard mask in the left/right (LR), superior/inferior (SI), and anterior/posterior (AP) translational planes, and Σ = 0.2 mm, 0.1 mm and 0.2 mm and σ = 0.2 mm, 0.3 mm and 0.2 mm, respectively, for the MFD. There was a reduction in rotation errors in the MFD compared to the mask. Margin calculations suggested an isotropic margin could be safely reduced to 2 mm for the MFD. CONCLUSION: The two devices demonstrate similar daily positional accuracy for fractionated stereotactic treatment of intracranial lesions. Combined with daily image guidance and couch correction, either of these devices is a viable frameless option for fractionated stereotactic radiation therapy.

Use of a megavoltage electronic portal imaging device to identify prosthetic materials.

To achieve accurate dose calculations in radiation therapy the electron density of patient tissues must be known. This information is ordinarily gained from a computed tomography (CT) image that has been calibrated to allow relative electron density (RED) to be determined from CT number. When high density objects such as metallic prostheses are involved, direct use of the CT data can become problematic due to the artefacts introduced by high attenuation of the beam. This requires manual correction of the density values, however the properties of the implanted prosthetic are not always known. A method is introduced where the RED of such an object can be determined using the treatment beam of a linear accelerator with an electronic portal imaging device. The technique was tested using a metallic hip replacement that was placed within a container of water. Compared to the theoretical RED of 6.8 for cobalt-chromium alloy, these measurements calculated a value of 6.4 ± 0.7. This would allow the distinction of an implant as Co-Cr or steel, which have similar RED, or titanium, which is much less dense with an RED of 3.7.

The dependence of computed tomography number to relative electron density conversion on phantom geometry and its impact on planned dose.

A computed tomography number to relative electron density (CT-RED) calibration is performed when commissioning a radiotherapy CT scanner by imaging a calibration phantom with inserts of specified RED and recording the CT number displayed. In this work, CT-RED calibrations were generated using several commercially available phantoms to observe the effect of phantom geometry on conversion to electron density and, ultimately, the dose calculation in a treatment planning system. Using an anthropomorphic phantom as a gold standard, the CT number of a material was found to depend strongly on the amount and type of scattering material surrounding the volume of interest, with the largest variation observed for the highest density material tested, cortical bone. Cortical bone gave a maximum CT number difference of 1,110 when a cylindrical insert of diameter 28 mm scanned free in air was compared to that in the form of a 30 × 30 cm(2) slab. The effect of using each CT-RED calibration on planned dose to a patient was quantified using a commercially available treatment planning system. When all calibrations were compared to the anthropomorphic calibration, the largest percentage dose difference was 4.2 % which occurred when the CT-RED calibration curve was acquired with heterogeneity inserts removed from the phantom and scanned free in air. The maximum dose difference observed between two dedicated CT-RED phantoms was ±2.1 %. A phantom that is to be used for CT-RED calibrations must have sufficient water equivalent scattering material surrounding the heterogeneous objects that are to be used for calibration.