Collimator optimization for small animal radiation therapy at a micro-CT.

PURPOSE: In radiation therapy of small animals treatment depths range from a few millimetres to several centimetres. In order to spare surrounding organs at risk steep dose gradients are necessary. To minimize the treatment time, and therefore the strain to the animals, a high dose rate is required. A description how these parameters can be optimized through an appropriate choice of collimators with different source surface distances (SSD) as well as different materials and geometries is presented. MATERIAL AND METHODS: An industrial micro-CT unit (Y.Fox, YXLON GmbH, Hamburg, Germany) was converted into a precision irradiator for small animals. Different collimators of either stainless steel (Fe) with cylindrical bores (SSD=42mm) or tungsten (W) with conical bores (SSD=14mm) were evaluated. The dosimetry of very small radiation fields presents a challenge and was performed with GafChromic EBT3 films (Ashland, Vayne, KY, USA) in a water phantom. The films were calibrated with an ionization chamber in the uncollimated field. Treatments were performed via a rotation of the objects with a fixed radiation source. RESULTS: As expected, the shorter SSD of the W-collimators resulted in a (4.5±1.6)-fold increase of the dose rates compared to the corresponding Fe-collimators. The ratios of the dose rates at 1mm and 10mm depth in the water phantom was (2.6±0.2) for the Fe- and (4.5±0.1) for the W-collimators. For rotational treatments in a cylindrical plastic phantom maximum dose rates of up to 1.2Gy/min for Fe- and 5.1Gy/min for W-collimators were measured. CONCLUSION: Choosing the smallest possible SSD leads to a high dose rate and a high surface dose, which is of advantage for the treatment of superficial target volumes. For larger SSD the dose rate is lower and the depth dose curve is shallower. This leads to a reduction of the surface dose and is best suited for treatments of deeper seated target volumes. Divergent collimator bores have, due to the reduced scatter within the collimators, a steeper penumbra. The dosimetry of small kilovoltage beams with Gafchromic EBT3 films in a water phantom has proven successful.

Imaging of Orthotopic Glioblastoma Xenografts in Mice Using a Clinical CT Scanner: Comparison with Micro-CT and Histology.

PURPOSE: There is an increasing need for small animal in vivo imaging in murine orthotopic glioma models. Because dedicated small animal scanners are not available ubiquitously, the applicability of a clinical CT scanner for visualization and measurement of intracerebrally growing glioma xenografts in living mice was validated. MATERIALS AND METHODS: 2.5x106 U87MG cells were orthotopically implanted in NOD/SCID/ᵞc-/- mice (n = 9). Mice underwent contrast-enhanced (300 µl Iomeprol i.v.) imaging using a micro-CT (80 kV, 75 µAs, 360° rotation, 1,000 projections, scan time 33 s, resolution 40 x 40 x 53 µm) and a clinical CT scanner (4-row multislice detector; 120 kV, 150 mAs, slice thickness 0.5 mm, feed rotation 0.5 mm, resolution 98 x 98 x 500 µm). Mice were sacrificed and the brain was worked up histologically. In all modalities tumor volume was measured by two independent readers. Contrast-to-noise ratio (CNR) and Signal-to-noise ratio (SNR) were measured from reconstructed CT-scans (0.5 mm slice thickness; n = 18). RESULTS: Tumor volumes (mean±SD mm3) were similar between both CT-modalities (micro-CT: 19.8±19.0, clinical CT: 19.8±18.8; Wilcoxon signed-rank test p = 0.813). Moreover, between reader analyses for each modality showed excellent agreement as demonstrated by correlation analysis (Spearman-Rho >0.9; p<0.01 for all correlations). Histologically measured tumor volumes (11.0±11.2) were significantly smaller due to shrinkage artifacts (p<0.05). CNR and SNR were 2.1±1.0 and 1.1±0.04 for micro-CT and 23.1±24.0 and 1.9±0.7 for the clinical CTscanner, respectively. CONCLUSION: Clinical CT scanners may reliably be used for in vivo imaging and volumetric analysis of brain tumor growth in mice.

The HIV-derived protein Vpr52-96 has anti-glioma activity in vitro and in vivo.

Patients with actively replicating human immunodeficiency virus (HIV) exhibit adverse reactions even to low irradiation doses. High levels of the virus-encoded viral protein R (Vpr) are believed to be one of the major underlying causes for increased radiosensitivity. As Vpr efficiently crosses the blood-brain barrier and accumulates in astrocytes, we examined its efficacy as a drug for treatment of glioblastoma multiforme (GBM).In vitro, four glioblastoma-derived cell lines with and without methylguanine-DNA methyltransferase (MGMT) overexpression (U251, U87, U251-MGMT, U87-MGMT) were exposed to Vpr, temozolomide (TMZ), conventional photon irradiation (2 to 6 Gy) or to combinations thereof. Vpr showed high rates of acute toxicities with median effective doses of 4.0±1.1 µM and 15.7±7.5 µM for U251 and U87 cells, respectively. Caspase assays revealed Vpr-induced apoptosis in U251, but not in U87 cells. Vpr also efficiently inhibited clonogenic survival in both U251 and U87 cells and acted additively with irradiation. In contrast to TMZ, Vpr acted independently of MGMT expression.Dose escalation in mice (n=12) was feasible and resulted in no evident renal or liver toxicity. Both, irradiation with 3x5 Gy (n=8) and treatment with Vpr (n=5) delayed intracerebral tumor growth and prolonged overall survival compared to untreated animals (n=5; p3x5 Gy<0.001 and pVpr=0.04; log-rank test).Our data show that the HIV-encoded peptide Vpr exhibits all properties of an effective chemotherapeutic drug and may be a useful agent in the treatment of GBM.

Image-Guided Radiotherapy Using a Modified Industrial Micro-CT for Preclinical Applications.

PURPOSE/OBJECTIVE: Although radiotherapy is a key component of cancer treatment, its implementation into pre-clinical in vivo models with relatively small target volumes is frequently omitted either due to technical complexity or expected side effects hampering long-term observational studies. We here demonstrate how an affordable industrial micro-CT can be converted into a small animal IGRT device at very low costs. We also demonstrate the proof of principle for the case of partial brain irradiation of mice carrying orthotopic glioblastoma implants. METHODS/MATERIALS: A commercially available micro-CT originally designed for non-destructive material analysis was used. It consists of a CNC manipulator, a transmission X-ray tube (10-160 kV) and a flat-panel detector, which was used together with custom-made steel collimators (1-5 mm aperture size). For radiation field characterization, an ionization chamber, water-equivalent slab phantoms and radiochromic films were used. A treatment planning tool was implemented using a C++ application. For proof of principle, NOD/SCID/γc(-/-) mice were orthotopically implanted with U87MG high-grade glioma cells and irradiated using the novel setup. RESULTS: The overall symmetry of the radiation field at 150 kV was 1.04 ± 0.02%. The flatness was 4.99 ± 0.63% and the penumbra widths were between 0.14 mm and 0.51 mm. The full width at half maximum (FWHM) ranged from 1.97 to 9.99 mm depending on the collimator aperture size. The dose depth curve along the central axis followed a typical shape of keV photons. Dose rates measured were 10.7 mGy/s in 1 mm and 7.6 mGy/s in 5 mm depth (5 mm collimator aperture size). Treatment of mice with a single dose of 10 Gy was tolerated well and resulted in central tumor necrosis consistent with therapeutic efficacy. CONCLUSION: A conventional industrial micro-CT can be easily modified to allow effective small animal IGRT even of critical target volumes such as the brain.

In vivo micro-CT imaging of untreated and irradiated orthotopic glioblastoma xenografts in mice: capabilities, limitations and a comparison with bioluminescence imaging.

Small animal imaging is of increasing relevance in biomedical research. Studies systematically assessing the diagnostic accuracy of contrast-enhanced in vivo micro-CT of orthotopic glioma xenografts in mice do not exist. NOD/SCID/γc(-/-) mice (n = 27) underwent intracerebral implantation of 2.5 × 10(6) GFP-Luciferase-transduced U87MG cells. Mice underwent bioluminescence imaging (BLI) to detect tumor growth and afterwards repeated contrast-enhanced (300 µl Iomeprol i.v.) micro-CT imaging (80 kV, 75 µAs, 360° rotation, 1,000 projections, 33 s scan time, resolution 40 × 40 × 53 µm, 0.5 Gy/scan). Presence of tumors, tumor diameter and tumor volume in micro-CT were rated by two independent readers. Results were compared with histological analyses. Six mice with tumors confirmed by micro-CT received fractionated irradiation (3 × 5 Gy every other day) using the micro-CT (5 mm pencil beam geometry). Repeated micro-CT scans were tolerated well. Tumor engraftment rate was 74 % (n = 20). In micro-CT, mean tumor volume was 30 ± 33 mm(3), and the smallest detectable tumor measured 360 × 620 µm. The inter-rater agreement (n = 51 micro-CT scans) for the item tumor yes/no was excellent (Spearman-Rho = 0.862, p < 0.001). Sensitivity and specificity of micro-CT were 0.95 and 0.71, respectively (PPV = 0.91, NPV = 0.83). BLI on day 21 after tumor implantation had a sensitivity and specificity of 0.90 and 1.0, respectively (PPV = 1.0, NPV = 0.5). Maximum tumor diameter and volume in micro-CT and histology correlated excellently (tumor diameter: 0.929, p < 0.001; tumor volume: 0.969, p < 0.001, n = 17). Irradiated animals showed a large central tumor necrosis. Longitudinal contrast enhanced micro-CT imaging of brain tumor growth in live mice is feasible at high sensitivity levels and with excellent inter-rater agreement and allows visualization of radiation effects.