Name: pet and mri guided irradiation of a glioblastoma rat model using a micro-irradiator
Uploaded: 2017-12-28T14:00:00
Description: Watch this Scientific Journal Video about PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator at JoVE.com

The authors would like to thank Stichting Luka Hemelaere and Soroptimist International for supporting this work.

The study was approved by the ethics committee for animal experiments (ECD 09/23 and ECD 12/28). All commercial details can be found in Table of Materials.
1. F98 GB Rat Cell Model
<ol>
	<li>Culture the F98 GB cells, obtained from ATCC, in monolayers using Dulbecco&#39;s modified Eagle Medium, 10% calf serum, 1% penicillin, 1% streptomycin, 1% L-glutamine, and 0.1% amphotericin b, and place in a CO2 incubator (5% CO2 and 37 &#176;C).</li>
	<li>Inoculate...

<ol>
	<li>Stupp, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Promising+survival+for+patients+with+newly+diagnosed+glioblastoma+multiforme+treated+with+concomitant+radiation+plus+temozolomide+followed+by+adjuvant+temozolomide.">Promising survival for patients with newly diagnosed glioblastoma multiforme treated with concomitant radiation plus temozolomide followed by adjuvant temozolomide.</a> J Clin Oncol. 20 (5), 1375-1382 (2002).</li><li>Dhermain, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiotherapy+of+high-grade+gliomas:+current+standards+and+new+concepts,+innovations+in+imaging+and+radiotherapy,+and+new+therapeutic+approaches.">Radiotherapy of high-grade gliomas: current standards and new concepts, innovations in imaging and radiotherapy, and new therapeutic approaches.</a> Chin J Cancer. 33 (1), 16-24 (2014).</li><li>Ahmed, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Malignant+gliomas:+current+perspectives+in+diagnosis,+treatment,+and+early+response+assessment+using+advanced+quantitative+imaging+methods.">Malignant gliomas: current perspectives in diagnosis, treatment, and early response assessment using advanced quantitative imaging methods.</a> Cancer Manag Res. 6, 149-170 (2014).</li><li>Verhaegen, F., Granton, P., Tryggestad, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+radiotherapy+research+platforms.">Small animal radiotherapy research platforms.</a> Phys Med Biol. 56 (12), R55-R83 (2011).</li><li>Kinsella, T. J., Vielhuber, K. A., Kunugi, K. A., Schupp, J., Davis, T. W., Sands, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+toxicity+and+efficacy+study+of+a+14-day+schedule+of+oral+5-iodo-2-pyrimidinone-2-deoxyribose+as+a+prodrug+for+5-iodo-2-deoxyuridine+radiosensitization+in+U251+human+glioblastoma+xenografts.">Preclinical toxicity and efficacy study of a 14-day schedule of oral 5-iodo-2-pyrimidinone-2-deoxyribose as a prodrug for 5-iodo-2-deoxyuridine radiosensitization in U251 human glioblastoma xenografts.</a> Clin Cancer Res. 6 (4), 1468-1475 (2000).</li><li>Vellimana, A. 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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+integrated+method+for+reproducible+and+accurate+image-guided+stereotactic+cranial+irradiation+of+brain+tumors+using+the+small+animal+radiation+research+platform.">An integrated method for reproducible and accurate image-guided stereotactic cranial irradiation of brain tumors using the small animal radiation research platform.</a> Transl Oncol. 5 (4), 230-237 (2012).</li><li>Grosu, A. -. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implications+of+IMT-SPECT+for+postoperative+radiotherapy+planning+in+patients+with+gliomas.">Implications of IMT-SPECT for postoperative radiotherapy planning in patients with gliomas.</a> Int J Radiat Oncol Biol Phys. 54 (3), 842-854 (2002).</li><li>Butterworth, K. T., Prise, K. M., Verhaegen, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+image-guided+radiotherapy:+Status,+considerations+and+potential+for+translational+impact.">Small animal image-guided radiotherapy: Status, considerations and potential for translational impact.</a> Br J Radiol. 88 (1045), 4-6 (2015).</li><li>Aird, E. G. A., Conway, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CT+simulation+for+radiotherapy+treatment+planning.">CT simulation for radiotherapy treatment planning.</a> Br J Radiol. 75 (900), 937-949 (2002).</li><li>Baker, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization:+Conventional+and+CT+simulation.">Localization: Conventional and CT simulation.</a> Br J Radiol. 79 (Spec No 1). , S36-S49 (2006).</li><li>Corroyer-Dumont, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI-guided+radiotherapy+of+the+SK-N-SH+neuroblastoma+xenograft+model+using+a+small+animal+radiation+research+platform.">MRI-guided radiotherapy of the SK-N-SH neuroblastoma xenograft model using a small animal radiation research platform.</a> Br J Radiol. 90 (1069), 20160427 (2017).</li><li>Bolcaen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI-guided+3D+conformal+arc+micro-irradiation+of+a+F98+glioblastoma+rat+model+using+the+Small+Animal+Radiation+Research+Platform+(SARRP).">MRI-guided 3D conformal arc micro-irradiation of a F98 glioblastoma rat model using the Small Animal Radiation Research Platform (SARRP).</a> J Neurooncol. 120 (2), 257-266 (2014).</li><li>Niyazi, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FET-PET+for+malignant+glioma+treatment+planning.">FET-PET for malignant glioma treatment planning.</a> Radiother Oncol. 99 (1), 44-48 (2011).</li><li>Grosu, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+experience+with+I-123-alpha-methyl-tyrosine+SPECT+in+the+3-D+radiation+treatment+planning+of+brain+gliomas.">First experience with I-123-alpha-methyl-tyrosine SPECT in the 3-D radiation treatment planning of brain gliomas.</a> Int J Radiat Oncol Biol Phys. 47 (2), 517-526 (2000).</li><li>Ling, C. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+multidimensional+radiotherapy+(MD-CRT):biological+imaging+and+biological+conformality.">Towards multidimensional radiotherapy (MD-CRT):biological imaging and biological conformality.</a> Int J Radiat Oncol Biol Phys. 47 (3), 551-560 (2000).</li><li>Wahl, R. L., Jacene, H., Kasamon, Y., Lodge, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+RECIST+to+PERCIST:+Evolving+Considerations+for+PET+response+criteria+in+solid+tumors.">From RECIST to PERCIST: Evolving Considerations for PET response criteria in solid tumors.</a> J Nucl Med. 50 (5), 122S-150S (2009).</li><li>Daisne, J. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+volume+in+pharyngolaryngeal+squamous+cell+carcinoma:+comparison+at+CT,+MR+imaging,+and+FDG+PET+and+validation+with+surgical+specimen.">Tumor volume in pharyngolaryngeal squamous cell carcinoma: comparison at CT, MR imaging, and FDG PET and validation with surgical specimen.</a> Radiology. 233 (1), 93-100 (2004).</li><li>Grosu, A. -. L., Weber, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PET+for+radiation+treatment+planning+of+brain+tumours.">PET for radiation treatment planning of brain tumours.</a> Radiother Oncol. 96 (3), 325-327 (2010).</li><li>Banissi, C., Ghiringhelli, F., Chen, L., Carpentier, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treg+depletion+with+a+low-dose+metronomic+temozolomide+regimen+in+a+rat+glioma+model.">Treg depletion with a low-dose metronomic temozolomide regimen in a rat glioma model.</a> Cancer Immunol Immunother. 58, 1627-1634 (2009).</li><li>Robinson, C. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+alternative+temozolomide+schedules+on+glioblastoma+O(6)-methylguanine-DNA+methyltransferase+activity+and+survival.">Effect of alternative temozolomide schedules on glioblastoma O(6)-methylguanine-DNA methyltransferase activity and survival.</a> Br J Cancer. 103, 498-504 (2010).</li><li>Espa&#241;a, S., Marcinkowski, R., Keereman, V., Vandenberghe, S., Van Holen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DigiPET:+sub-millimeter+spatial+resolution+small-animal+PET+imaging+using+thin+monolithic+scintillators.">DigiPET: sub-millimeter spatial resolution small-animal PET imaging using thin monolithic scintillators.</a> Phys Med Biol. 59 (13), 3405 (2014).</li></ol>

To achieve accurate irradiation of the glioblastoma tumor target in the rat brain, the micro-irradiator&#39;s on-board CT guidance was not sufficient. Brain tumors are hardly visible due to insufficient soft tissue contrast, even if contrast enhancement would be used. As such, MRI needs to be included to allow more precise irradiation. Using a sequential MR acquisition on a 7 T system and a CT acquisition on the micro-irradiator we were able to target the dose to the contrast-enhancing tumor tissue in the brain and calcu...

High-grade glioma is the most common and most aggressive malignant brain tumor in adults with a median survival of 1 year despite current treatment modalities. The standard of care includes maximal surgical resection followed by combined external beam radiation therapy (RT) and temozolomide (TMZ), followed by maintenance TMZ1,2,3. Since the introduction of TMZ now more than 15 years ago, no significant improvements have been made in the treatment of these tumors. Therefore, the implementation of new therapeutic strategies is urgent but should be first investigated in small animal cancer therapy models (mostly mice and rats). Tumor-bearing rodent models can be used to investigate the efficacy of new and complex radiation protocols, possibly combined with other (new) treatment agents, to assess radiation response or to investigate radio-protective agents. A major advantage of preclinical radiation research is the ability to work under controlled experimental conditions using large cohorts resulting in accelerated data yield due to the shorter lifespans of rodents. The preclinical findings should then be translated into a clinical trial in a much faster and more efficient way than in current practice4.
Small animal radiation experiments in the last decades have typically been achieved using fixed radiation sources5,6,7, e.g., 137Cs and 60Co, isotopes, or linear accelerators intended for human clinical use, applying a single radiation field with MV X-rays6,8,9,10,11. However, these devices do not reach sub-millimeter precision, which is required for small animals12. Furthermore, MV X-rays have characteristics unsuitable for irradiating small targets, such as a dose build-up at the air-tissue interface in the entrance region of the beam with an extent in the order of the animal size itself4,6,8,9,10,11. The latter makes it quite challenging to deliver a uniform dose to a tumor while sparing surrounding normal brain tissue4,8,9,10,11. Hence, it is unclear to which extent current animal studies still are relevant for modern RT practice12. In this respect, recently developed three-dimensional (3D) conformal small animal micro-irradiators are promising to bridge the technological gap between advanced 3D image-guided RT techniques, such as intensity modulated radiation therapy (IMRT) or conformal arcs used in humans and current small animal irradiation4,13. These platforms make use of a kilovoltage (kV) X-ray source to obtain sharp penumbras and to avoid dose build-up. These platforms include a computer-controlled stage for animal positioning, a kV X-ray source for imaging and radiation treatment, a rotational gantry assembly to allow radiation delivery from various angles, and a collimating system to shape the radiation beam4. In 2011, a micro-irradiator was installed at the preclinical imaging lab of Ghent University (Figure 1). This system is similar to modern human radiotherapy practice and enables a wide variety of preclinical experiments, such as the synergy of radiation with other therapies, complex radiation schemes, and image-guided sub-target boost studies.
Treatment planning on these micro-irradiators is based on CT, which is equivalent to human planning systems14,15. For CT imaging, an on-board X-ray detector is used in combination with the same kV X-ray tube that is used during treatment. CT imaging is used as it allows for accurate animal positioning and provides information necessary for individual radiation dose calculations via segmentation. However, due to the low soft-tissue contrast in CT imaging, tumors in the brain of small animals, such as high-grade glioma, cannot be easily delineated. The incorporation of multi-modality imaging is therefore necessary for an accurate target volume delineation. Compared to CT, MRI provides vastly superior soft-tissue contrast. This makes it much easier to visualize lesion boundaries that will result in a much better delineation of the target volume, helping to better irradiate the lesion and avoid surrounding tissue, as illustrated in Figure 24,16. An additional advantage is that MRI uses non-ionizing radiation, unlike CT that is using ionizing radiation. The major disadvantages of MRI are the relatively long acquisition times and high operational costs. It is important to note that MRI scans cannot be used for dose calculations, as they do not provide the required electron density information, although progress is being made in this field, too with the recent development of MR-LINACS. As such, a combined CT/MRI dataset is the method of choice for planning the irradiation of malignant glioma, containing both the information required for targeting (MRI-based volumes) and for dose calculations (CT-based electron density).
To decrease the gap between small animal irradiation and clinical routine, MRI clearly needs to be integrated into the work flow of the micro-irradiator, requiring a correct registration between MRI and CT, which is far from trivial. In this paper, our protocol for MRI-guided 3D conformal irradiation of F98 glioblastoma in rats is discussed, which has been published recently17.
Although incorporating CT and MRI in the workflow of the micro-irradiator is a clear step forward in small animal irradiation research, these anatomical imaging techniques do not always allow a full definition of the target volume. Pathological changes in the brain on CT and MRI are characterized by increased water content (edema) and leakage of the blood-brain barrier or contrast enhancement. However, both contrast-enhancement and hyper-intense areas on T2-weighted MRI are not always an accurate measure of tumor extent. Tumor cells have been detected far beyond the margins of contrast-enhancement12. Also, none of these techniques can identify the most aggressive parts within the tumor, which may be responsible for therapeutic resistance and tumor recurrence. Therefore, additional information from molecular imaging techniques like PET may have an added value for RT target volume definition because these techniques enable to visualize biologic pathways in vivo12,18,19.
In 2000, Ling et al. introduced the concept of biological target volume (BTV) by integrating anatomical and functional imaging into the radiotherapy workflow, leading to what they called multidimensional conformal radiotherapy20. This creates the possibility to improve dose targeting by delivering a non-uniform dose to a target region using for example PET images. The most widely used PET tracer for tumor staging and to monitor treatment response is fluor-18 (18F) labeled fluorodeoxyglucose (FDG), which visualizes the glucose metabolism21. In head and neck cancer, previous studies have shown that the use of 18F-FDG PET led to a better estimate of the actual tumor volume, as defined by the pathologic specimens, compared with CT and MRI22. In primary brain tumors, where FDG is not useful due to the very strong background signal from the normal brain, amino acids, such as 11C-methionine and more recently 18F-fluoroetthyltyrosine (FET), have been investigated for GTV delineation with often marked differences between amino-acid PET and MRI-based GTVs23. However, no prospective trial investigating the meaning of this finding has been performed yet. In this study, we selected the amino-acid tracer 18F-FET and the hypoxia tracer 18F-fluoroazomycin-arabinoside (18F-FAZA). 18F-FET and 18F-FAZA were selected because an increased amino-acid uptake is strongly correlated with the proliferation rate in GB tumors, whereas uptake of a hypoxia PET-tracer is correlated with resistance to (chemo)radiotherapy18,23. Sub-volume boosting using the micro-irradiator was optimized by giving an additional radiation dose to a PET-defined part of the F98 GB tumor in rats.

<table>
	<tbody>
		<tr>
			<td>GB RAT model</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>F98 Glioblastoma cell line</td>
			<td>ATCC</td>
			<td>CRL-2397</td>
			<td></td>
		</tr>
		<tr>
			<td>Fischer F344/Ico crl Rats</td>
			<td>Charles River</td>
			<td>N/A</td>
			<td>http://www.criver.com/products-services/basic-research/find-a-model/fischer-344-rat</td>
		</tr>
		<tr>
			<td>Micropump system</td>
			<td>World Precision Instruments</td>
			<td>UMP3</td>
			<td>Micro 4: https://www.wpiinc.com/products/top-products/make-selection-ump3-ultramicropump/#tabs-1</td>
		</tr>
		<tr>
			<td>Stereotactic frame</td>
			<td>Kopf</td>
			<td>902</td>
			<td>Model 902 Dual Small Animal Stereotaxic frame</td>
		</tr>
		<tr>
			<td>diamant drill</td>
			<td>Velleman</td>
			<td>VTHD02</td>
			<td>https://www.velleman.eu/products/view/?id=370450</td>
		</tr>
		<tr>
			<td>Bone wax</td>
			<td>Aesculap</td>
			<td>1029754</td>
			<td>https://www.aesculapusa.com/products/wound-closure/hemostatic-bone-wax</td>
		</tr>
		<tr>
			<td>Insulin syringe Microfine</td>
			<td>Beckton-Dickinson</td>
			<td>320924</td>
			<td>1 mL, 29G</td>
		</tr>
		<tr>
			<td>InfraPhil IR lamp</td>
			<td>Philips</td>
			<td>HP3616/01</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethilon</td>
			<td>Ethicon</td>
			<td>662G/662H</td>
			<td>FS-2, 4-0, 3/8, 19 mm</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Cell culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Invitrogen</td>
			<td>14040-091</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicilline-streptomycine</td>
			<td>Invitrogen</td>
			<td>15140-148</td>
			<td></td>
		</tr>
		<tr>
			<td>L-glutamine</td>
			<td>Invitrogen</td>
			<td>25030-032</td>
			<td></td>
		</tr>
		<tr>
			<td>Fungizone</td>
			<td>Invitrogen</td>
			<td>15290-018</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin-EDTA</td>
			<td>Invitrogen</td>
			<td>25300-062</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Invitrogen</td>
			<td>14040-224</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcons</td>
			<td>Thermo Scientific</td>
			<td>178883</td>
			<td>175 cm2 nunclon surface, disposables for cell culture with filter caps</td>
		</tr>
		<tr>
			<td>Cell freezing medium</td>
			<td>Sigma-aldrich</td>
			<td>C6164</td>
			<td>Cell Freezing Medium-DMSO, sterile-filtered, suitable for cell culture, endotoxin tested</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Animal irradiation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-irradiator</td>
			<td>X-strahl</td>
			<td>SARRP</td>
			<td></td>
		</tr>
		<tr>
			<td>software for irradiation</td>
			<td>X-strahl</td>
			<td>MuriPlan</td>
			<td>pre-clinical treatment planning system (PCTPS), version 2.0.5.</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Small animal PET</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>microPET system possibility 1</td>
			<td>Molecubes</td>
			<td>B-Cube</td>
			<td>http://www.molecubes.com/b-cube/</td>
		</tr>
		<tr>
			<td>microPET system possibility 2</td>
			<td>TriFoil Imaging, Northridge CA</td>
			<td>FLEX Triumph II</td>
			<td>http://www.trifoilimaging.com</td>
		</tr>
		<tr>
			<td>PET tracers</td>
			<td>In-house made</td>
			<td></td>
			<td>18F-FDG, 18F-FET, 18F-FAZA, 18F-Choline</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Small animal MRI</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>microMRI system</td>
			<td>Bruker Biospin</td>
			<td>Pharmascan 70/16</td>
			<td>https://www.bruker.com/products/mr/preclinical-mri/pharmascan/overview.html</td>
		</tr>
		<tr>
			<td>Dotarem contrast agent</td>
			<td>Guerbet</td>
			<td></td>
			<td>MRI contrast agent, Dotarem 0,5 mmol/ml</td>
		</tr>
		<tr>
			<td>rat whole body transmitter coil</td>
			<td>Rapid Biomedical</td>
			<td>V-HLS-070</td>
			<td></td>
		</tr>
		<tr>
			<td>rat brain surface coil</td>
			<td>Rapid Biomedical</td>
			<td>P-H02LE-070</td>
			<td></td>
		</tr>
		<tr>
			<td>Water-based heating unit</td>
			<td>Bruker Biospin</td>
			<td>MT0125</td>
			<td></td>
		</tr>
		<tr>
			<td>30 G Needle for IV injection</td>
			<td>Beckton-Dickinson</td>
			<td>305128</td>
			<td>30 G</td>
		</tr>
		<tr>
			<td>PE 10 tubing (60 cm/injection)</td>
			<td>Instech laboratories, Inc</td>
			<td>BTPE-10</td>
			<td>BTPE-10, polyethylene tubing 0.011 x .024 in (0.28 x 60 mm), non sterile, 30 m (98 ft) spool, Instech laboratories, Inc Plymouth meeting PA USA- (800) 443-4227- http://www.instechlabs.com</td>
		</tr>
		<tr>
			<td>non-heparinised micro haematocrit capillaries</td>
			<td>GMBH</td>
			<td>7493 21</td>
			<td>these capillaries are filled with water to create markers visible on MRI and CT</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Consumables</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>isoflurane: Isoflo</td>
			<td>Zoetis</td>
			<td>B506</td>
			<td>Anaesthesia</td>
		</tr>
		<tr>
			<td>ketamine: Ketamidor</td>
			<td>Ecuphar</td>
			<td></td>
			<td>Anaesthesia</td>
		</tr>
		<tr>
			<td>xylazine: Sedaxyl</td>
			<td>Codifar NV</td>
			<td></td>
			<td>Anaesthesia</td>
		</tr>
		<tr>
			<td>catheter</td>
			<td>Terumo</td>
			<td>Versatus-W</td>
			<td>26G</td>
		</tr>
		<tr>
			<td>Temozolomide</td>
			<td>Sigma-aldrich</td>
			<td>T2577-100MG</td>
			<td>chemotherapy</td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma-aldrich</td>
			<td>276855-100ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringe Microfine</td>
			<td>Beckton-Dickinson</td>
			<td>320924</td>
			<td>1 mL, 29G</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Image analysis</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PMOD software</td>
			<td>PMOD technologies LLC</td>
			<td>PFUS (fusion tool)</td>
			<td>biomedical image quantification software (BIQS), version 3.405, https://www.pmod.com/web/?portfolio=22-image-processing-pfus</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Anesthesia-equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthetic movabe unit</td>
			<td>ASA LTD</td>
			<td>ASA 0039</td>
			<td>ASA LTD, 5 valley road, Keighley, BD21 4LZ</td>
		</tr>
		<tr>
			<td>Oxygen generator</td>
			<td>Veterinary technics Int.</td>
			<td>7F-3</td>
			<td>BDO-Medipass, Ijmuiden</td>
		</tr>
	</tbody>
</table>

pet and mri guided irradiation of a glioblastoma rat model using a micro-irradiator

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without the ability to target a specific or well-delineated tumor volume. The delivery of radiation was achieved using fixed radiation sources or linear accelerators producing megavoltage (MV) X-rays. These devices are unable to achieve sub-millimeter precision required for small animals. Furthermore, the high doses delivered to healthy surrounding tissue hamper response assessment. To increase the translation between small animal studies and humans, our goal was to mimic the treatment of human glioblastoma in a rat model. To enable a more accurate irradiation in a preclinical setting, recently, precision image-guided small animal radiation research platforms were developed. Similar to human planning systems, treatment planning on these micro-irradiators is based on computed tomography (CT). However, low soft-tissue contrast on CT makes it very challenging to localize targets in certain tissues, such as the brain. Therefore, incorporating magnetic resonance imaging (MRI), which has excellent soft-tissue contrast compared to CT, would enable a more precise delineation of the target for irradiation. In the last decade also biological imaging techniques, such as positron emission tomography (PET) gained interest for radiation therapy treatment guidance. PET enables the visualization of e.g., glucose consumption, amino-acid transport, or hypoxia, present in the tumor. Targeting those highly proliferative or radio-resistant parts of the tumor with a higher dose could give a survival benefit. This hypothesis led to the introduction of the biological tumor volume (BTV), besides the conventional gross target volume (GTV), clinical target volume (CTV), and planned target volume (PTV).
At the preclinical imaging lab of Ghent University, a micro-irradiator, a small animal PET, and a 7 T small animal MRI are available. The goal was to incorporate MRI-guided irradiation and PET-guided sub-volume boosting in a glioblastoma rat model.

To mimic the human treatment methodology for the irradiation of glioblastoma in a preclinical model, inclusion of MRI-guided radiotherapy was necessary. Using the PCTPS and the micro-irradiator interface we were able to irradiate F98 glioblastoma in rats with multiple conformal non-coplanar arcs targeting the contrast-enhanced region on T1-weighted MRI17. Rigid-body transformations in combination with a multi-modality bed were used for image registration between MR...

Watch this Scientific Journal Video about PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator at JoVE.com

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without ...

The overall goal of this methodology is to perform image-guided conformal irradiation in small animals. This method can help to answer key questions in the radiology field about how to delineate specific tumor volumes for targeted radiation therapy. The main advantage of this technique is that it mimics the human treatment of cancer and allows for cells irradiation of tumors in rats.Using PETs to guide to deradiation beams, allows to take into account cancer biology as a new and promising development in the field of small animal radiotherapy. The inclusion of a biological target volume, allows the most active and radiation resistant areas of the tumors to be targeted, resulting in better outcomes of the therapy. To inoculate the brain of an anesthetized 170 gram female fisher 5344 rat with glioma cells, female fisher 5344 rat with glioma cells, first, confirm sedation by lack of response to toe pinch, remove the hair from eye level to the back of the skull, and apply ointment to the animal's eyes.Immobilize the animal in a stereotactic device. Disinfect the exposed skin with povidone iodine, and expose the skull with a two centimeter midline scalp incision. Using a diamond drill, make a one millimeter hole two millimeters posterior and two and a half millimeters lateral to the bregma in the right frontal hemisphere.Next, load the needle of a 29 gauge insulin syringe with five microliters of cell suspension, use a microsyringe pump controller to inject the cells three millimeters deep into the skull under stereotactic guidance and withdraw the needle slowly. Close the incision with bone wax. Then, suture and disinfect the skin with more povidone iodine and use a red lamp to stabilize the body temperature of the animal post surgery with monitoring until full recovery.Eight days post inoculation, connect a 30 gauge needle to a 60 centimeter long tube. Positioned intravenously within the lateral tail vein, and place the anesthetized animal into an MRI bed. Place the bed in the holder with a fixed rat brain surface coil, and position the bed in a 72 millimeter rat hold body transmitor coil.Then, assess the tumor growth with a localizor scan, followed by a T2 weighted spin echo scan. If a tumor is confirmed, start a 12 minute dynamic contract enhanced MRI acquisition, injecting a gadolinium containing contrast agent into the intraveniously placed tubing 30 seconds after beginning the scan. To plot the signal intensity over time, use the image sequence analysis tool to select a region of interest within the suspected tumor region and analyze the shape of the resulting dynamic contract enhanced curve to confirm the presence of the glioblastoma.Then, acquire a contrast enhanced T1 weighted, spin echo sequence. For multi-modality imaging of the target volume, insert a 26 gauge catheter into the tail vein and inject 37 megabecquerels of the PET radioactive tracer of interest and 200 microliters of saline into the catheter. 15 minutes before the PET acquisition, inject the MRI contrast agent through the tail vein catheter and place the anesthetized rat on a custom made multi-modality bed.Position one multi-modality marker under, above, and on the right side of the skull. Using hook and loop fasteners, secure the rat to the bed. Place the bed in the animal holder of the MRI scanner, fix the rat brain surface coil, and position the entire setup in the 72 millimeter rat hold body transmitor coil.Obtain a localizer scan followed by a contrast enhanced T1 weighted spin echo sequence as demonstrated. At the end of the T1 scan, transfer the animal to the PET instrument and obtain the appropriate 30 minute static PET scan in list mode according to the parameters for the injected PET tracer. Then, transfer the bed onto a plastic holder secured onto the four axis robotic positioning table of the micro-irradiator.And obtain a high resolution treatment planning CT scan using an aluminum filter of one millimeter and a 20 by 20 centimeter amorphous silicon flat panel detector. Select manually gray value thresholds until you achieve a good segmentation of bone, soft tissue and air. Make sure that there is no air inside the skull.For treatment planning, import the planning computed tomography, or CT, into the pre-clinical treatment planning system, or PCTPS, and manually segment the CT image into three different tissue classes. A precise fusion can be achieved by over laying the increased signal intensity of the skull on the CT scan with the black signal on the MRI scan. Load MRI scan and co-register using rigid transformations and the multi-modality markers and the skull.Load the MRI into the PCTPS. Then first fill in the transformation matrix. Switch from CT to MRI and back to check the fusion and add left, right, posterior, anterior, and inferior, superior transformations and rotations until the perfect fusion is achieved.Then, select the target or irradiation in the center of the contrast enhancing tumor on the T1 weighted MRI. If additional PET information must be included, use the biomedical image quantification software to include a CT/MRI PETCO registration. First, load the CT scan, afterwards load the PET scan.Check the orientation of the PET scan when loading. Change the color scale of the PET image, and the orientation. Apply a gaussian filter to the image, such that the tracer uptake in the tumor becomes clearly visible.Adjust the CT contrast to start the image fusion process to achieve a PET MRI image fusion, and use the contouring tool in the quantification software. After co-registration, select the target in the center of the increased PET tracer uptake in the quantification software. Use both rotations and translations and check the fusion in all the slices of the image.Select the center of the region with the highest tracer uptake and extract the coordinates. And manually enter the coordinates into the PCTPS. If the automatic PET MRI and CT image fusion tools do not generate a good fusion, contouring tools and manually transformations can be used to improve the fusion results.Select the prescribed dose, number of arcs, arc position, rotation range of the arcs, and the collimator size and adjust the settings for the appropriate MRI or PET-MRI guided radiation therapy. For the actually irradiation, select a 0.5 millimeter copper filter, set the x-ray voltage to 220 kilovolts, and the x-ray current to 13 milliamps, and position the right collimator on the gantry. Then, transfer the appropriate beam delivery parameters from the PCTPS to the micro-irradiator to execute the radiation therapy.To mimic the human treatment methodology for glioblastoma irradiation in a pre-clinical model, the iso-center for irradiation is selected in the center of the contrast enhanced tumor region on the T1 weighted MRI, as just demonstrated. In this experiment, distributions and cumulative dose volume histograms of the mean, minimal and maximal doses of the target volume, and the normal brain tissue volumes, were calculated for five different animals. For co-registration of the MRI, and CT modalities, the bio-medical image quantification software enables the use of many tools for rigid matching.By applying a simple transformation, both the MRI and PET based iso-centers can be transferred to the PCTPS for dose calculation of the radiation within each iso-center. One small certain technique can be use to irradiate tumors in rats and mice in lesson to our if performed properly. By performing this procedure, it's important to remember to monitor the animals carefully while they are under anesthesia.After watching this video, you should have a good understanding of how to apply image guided, small animal irradiation of tumor targets of interest.

pet-mri-guided-irradiation-glioblastoma-rat-model-using-micro

Research

JoVE Journal

Cancer Research

Endobronchial Ultrasound-guided Intratumoral Injection of Cisplatin for the Treatment of Isolated Mediastinal Recurrence of Lung Cancer

Isolated hilar and mediastinal recurrence (IMHR) following external beam radiation therapy (EBRT) in patients with lung cancer is common. These patients do not have many treatment options and are usually offered palliative chemotherapy or best supportive care. Endobronchial ultrasound (EBUS)-guided intratumoral injection of cisplatin (ITC) is a novel approach for these patients. The procedure is performed under conscious sedation. The lesion is located with a bronchoscopy using EBUS, and a 22-gauge EBUS needle is advanced through the working channel of the scope and locked in position. Under ultrasound guidance, the wall of the tracheobronchial tree is punctured and the needle is moved into the target lesion. The needle stylet is then removed and cisplatin (40 mg/40 mL) is injected into the lesion. One to two sites are treated per session. Details of the procedure are described in the protocol section of paper. At our center, 50 sites were treated in 36 patients (19 males, 17 females). The mean age of our cohort was 61.9 &#177;8.5 years. We performed final analyses on 35 patients and 41 sites. 24/35 (69%) had complete or partial response (responders), whereas 11/35 (31%) had stable or progressive disease (non-responders). Overall, survival in our group was 8 months (95% CI of 6-11 months), with patients who responded having significantly better survival than the ones who did not.

The management of isolated recurrent lung cancer in a previously-irradiated field is challenging. Here, we describe an endobronchial ultrasound (EBUS)-guided cisplatin injection for the management of patients with localized lung cancer recurrence in a previously-radiated field.

Isolated hilar and mediastinal recurrence (IMHR) following external beam radiation therapy (EBRT) in patients with lung cancer is common. These patients ...

A Mouse Model of Fatigue Induced by Peripheral Irradiation

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. Here we describe a method using targeted peripheral irradiation to induce fatigue-like behavior in mice. With appropriate shielding, the irradiation targets the lower abdominal/pelvic region of the mouse, sparing the brain, in an effort to model radiation treatment received by individuals with pelvic cancers. We deliver an irradiation dose that is sufficient to induce fatigue-like behavior in mice, measured by voluntary wheel-running activity (VWRA), without causing obvious morbidity. Since wheel running is a normal, voluntary behavior in mice, its use should have little confounding effect on other behavioral tests or biological measures. Hence, wheel running can be used as a feasible outcome measure in understanding the behavioral and biological correlates of fatigue. CRF is a complex condition with frequent comorbidities, and likely has causes related both to cancer and its various treatments. The methods described in this paper are useful for investigating radiation-induced changes that contribute to the development of CRF and, more generally, to explore the biological networks that can explain the development and persistence of a peripherally-triggered but centrally-driven behavior like fatigue.

We describe a method using targeted peripheral irradiation to induce fatigue-like behavior in mice. The selected non-lethal irradiation dose leads to a week-long reduction in voluntary wheel-running activity.

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. ...

Whole-body PET/MRI of Pediatric Patients: The Details That Matter

Integrated PET/MRI is a hybrid imaging technique enabling clinicians to acquire diagnostic images for tumor assessment and treatment monitoring with both high soft tissue contrast and added metabolic information. Integrated PET/MRI has shown to be valuable in the clinical setting and has many promising future applications. The protocol presented here will provide step-by-step instructions for the acquisition of whole-body 2-deoxy-2-(18F)fluoro-D-glucose (18F-FDG) PET/MRI data in children with cancer. It also provides instructions on how to combine a whole-body staging scan with a local tumor scan for evaluation of the primary tumor. The focus of this protocol is to be both comprehensive and time-efficient, which are two ubiquitous needs for clinical applications. This protocol was originally developed for children above 6 years, or old enough to comply with breath-hold instructions, but can also be applied to patients under general anesthesia. Similarly, this protocol can be modified to fit institutional preferences in terms of choice of MRI pulse sequences for both the whole-body scan and local tumor assessment.

This article provides comprehensive step-by-step instructions for the acquisition of whole-body 2-deoxy-2-(18F)fluoro-D-glucose (18F-FDG) PET/MRI scans for cancer staging of pediatric patients. The protocol was developed for children above 6 years, or old enough to comply with breath-hold instructions, but can be used for general anesthesia patients as well.

Integrated PET/MRI is a hybrid imaging technique enabling clinicians to acquire diagnostic images for tumor assessment and treatment monitoring with both ...

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting immunodeficient mice subcutaneously with cancer cells and measuring the tumor mass after it becomes visible and palpable. Orthotopic injections of cancer cells aim to introduce the xenograft in the microenvironment that most closely resembles the tissue of origin of the tumor being studied. Brain cancer research requires intracranial injection of cancer cells to allow the tumor formation and analysis in the unique microenvironment of the brain. The in vivo imaging of intracranial xenografts monitors instantaneously the tumor mass of orthotopically engrafted mice. Here we report the use of fluorescence molecular tomography (FMT) of brain tumor xenografts. The cancer cells are first transduced with near infrared fluorescent proteins and then injected in the brain of immunocompromised mice. The animals are then scanned to obtain quantitative information about the tumor mass over an extended period of time. Cell pre-labeling allows for cost effective, reproducible, and reliable quantification of the tumor burden within each mouse. We eliminated the need for injecting imaging substrates, and thus reduced the stress on the animals. A limitation of this approach is represented by the inability to detect very small masses; however, it has better resolution for larger masses than other techniques. It can be applied to evaluate the efficacy of a drug treatment or genetic alterations of glioma cell lines and patient-derived samples.

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Orthotopic intracranial injection of tumor cells has been used in cancer research to study brain tumor biology, progression, evolution, and therapeutic response. Here we present fluorescence molecular tomography of tumor xenografts, which provides real-time intravital imaging and quantification of a tumor mass in preclinical glioblastoma models.

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting ...

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. Although heterogeneous in presentation, a common denominator among these tumors is the overexpression of somatostatin receptors. 68Ga-DOTATATE is a somatostatin analog labeled with the positron emitter gallium-68 (68Ga). For well-differentiated neuroendocrine tumors, 68Ga-DOTATATE positron emission tomography (PET)/computed tomography (CT) imaging is used for diagnosis, determination of disease burden, and therapy selection.
This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiolabeling of 68Ga-DOTATATE is performed with a fully automated labeling module coupled to a germanium-68 (68Ge)/68Ga generator. Quality control of the final product evaluates radiochemical purity with instant thin-layer chromatography and solid-phase chromatography, and pH prior to patient injection. Periodic quality control is performed to determine 68Ge breakthrough, sterility, and (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) content. Patient preparation includes patient instructions, a protocol for 68Ga-DOTATATE during treatment with somatostatin analogs, and intravenous administration of the radiopharmaceutical. For PET/CT imaging, the acquisition and reconstruction settings are described. For each step, radiation safety will be highlighted, as well as time constrictions due to the short half-life of 68Ga.
Fully automated in-house production and quality control of 68Ga-DOTATATE leads to very high success rates (95%) and produces two to four patient dosages per batch, depending on the yield of the generator. In conclusion, 68Ga-DOTATATE PET/CT imaging is a noninvasive and fast method of providing information on the tumor burden of neuroendocrine tumors (NETs) while also assisting in diagnosis and therapy selection.

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Well-differentiated neuroendocrine tumors overexpress somatostatin receptors which can be utilized for diagnostic imaging with the radiolabeled somatostatin analog 68Ga-DOTATATE. This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiation safety and time constrictions due to the short half-life of 68Ga are taken into account.

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less aggressive breast tumors, the 5-year survival rate of TNBC patients is 77% due to their characteristic drug-resistant phenotype and metastatic burden. Toward this end, murine models have been established aimed at identifying novel therapeutic strategies limiting TNBC tumor growth and metastatic spread. This work describes a practical guide for the TNBC orthotopic model where MDA-MB-231 breast cancer cells suspended in a basement membrane matrix are implanted in the fourth mammary fat pad, which closely mimics the cancer cell behavior in humans. Measurement of tumors by caliper, lung metastasis assessment via in vivo and ex vivo imaging, and molecular detection are discussed. This model provides an excellent platform to study therapeutic efficacy and is especially suitable for the study of the interaction between the primary tumor and distal metastatic sites.

This work presets an advanced protocol to accurately assess tumor loading by detection of green fluorescent protein and bioluminescence signals as well as the integration of quantitative molecular detection technique.

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

A 3D Spheroid Model for Glioblastoma

Two-dimensional (2D) cell cultures do not mimic in vivo tumor growth satisfactorily. Therefore, three-dimensional (3D) culture spheroid models were developed. These models may be particularly important in the field of neuro-oncology. Indeed, brain tumors have the tendency to invade the healthy brain environment. We describe herein an ideal 3D glioblastoma spheroid-based assay that we developed to study tumor invasion. We provide all technical details and analytical tools to successfully perform this assay.

Here, we describe an easy-to-use invasion assay for glioblastoma. This assay is suitable for glioblastoma stem-like cells. A Fiji macro for easy quantification of invasion, migration, and proliferation is also described.

Two-dimensional (2D) cell cultures do not mimic in vivo tumor growth satisfactorily. Therefore, three-dimensional (3D) culture spheroid models were ...

A Syngeneic Orthotopic Osteosarcoma Sprague Dawley Rat Model with Amputation to Control Metastasis Rate

The most recent advance in the treatment of osteosarcoma (OS) occurred in the 1980s when multi-agent chemotherapy was shown to improve overall survival compared to surgery alone. To address this problem, the aim of the study is to refine a lesser-known model of OS in rats with a comprehensive histologic, imaging, biologic, implantation, and amputation surgical approach that prolongs survival. We used an immunocompetent, outbred Sprague-Dawley (SD), syngeneic rat model with implanted UMR106 OS cell line (originating from a SD rat) with orthotopic tibial tumor implants into 3-week-old male and female rats to model pediatric OS. We found that rats develop reproducible primary and metastatic pulmonary tumors, and that limb amputations at 3 weeks post implantation significantly reduce the incidence of pulmonary metastasis and prevent unexpected deaths. Histologically, the primary and metastatic OSs in rats were very similar to human OS. Using immunohistochemistry methods, the study shows that rat OS are infiltrated with macrophages and T cells. A protein expression survey of OS cells reveals that these tumors express ErbB family kinases. Since these kinases are also highly expressed in most human OSs, this rat model could be used to test ErbB pathway inhibitors for therapy.

Here, a syngeneic orthotopic implantation followed by an amputation procedure of the osteosarcoma with spontaneous pulmonary metastasis that can be used for preclinical investigation of metastasis biology and development of novel therapeutics is described.

The most recent advance in the treatment of osteosarcoma (OS) occurred in the 1980s when multi-agent chemotherapy was shown to improve overall survival ...

Positron Emission Tomography-based Dose Painting Radiation Therapy in a Glioblastoma Rat Model using the Small Animal Radiation Research Platform

A rat glioblastoma model to mimic chemo-radiation treatment of human glioblastoma in the clinic was previously established. Similar to the clinical treatment, computed tomography (CT) and magnetic resonance imaging (MRI) were combined during the treatment-planning process. Positron emission tomography (PET) imaging was subsequently added to implement sub-volume boosting using a micro-irradiation system. However, combining three imaging modalities (CT, MRI, and PET) using a micro-irradiation system proved to be labor-intensive because multimodal imaging, treatment planning, and dose delivery have to be completed sequentially in the preclinical setting. This also results in a workflow that is more prone to human error. Therefore, a user-friendly algorithm to further optimize preclinical multimodal imaging-based radiation treatment planning was implemented. This software tool was used to evaluate the accuracy and efficiency of dose painting radiation therapy with micro-irradiation by using an in silico study design. The new methodology for dose painting radiation therapy is superior to the previously described method in terms of accuracy, time efficiency, and intra- and inter-user variability. It is also an important step towards the implementation of inverse treatment planning on micro-irradiators, where forward planning is still commonly used, in contrast to clinical systems.

Here we present a protocol to perform preclinical positron emission tomography-based radiotherapy in a rat glioblastoma model using algorithms developed in-house to optimize the accuracy and efficiency.

A rat glioblastoma model to mimic chemo-radiation treatment of human glioblastoma in the clinic was previously established. Similar to the clinical ...

Magnetic Resonance-Guided High Intensity Focused Ultrasound Generated Hyperthermia: A Feasible Treatment Method in a Murine Rhabdomyosarcoma Model

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time imaging and acoustic energy modulation, this modality enables precise temperature control within a defined area. Many thermal applications are being explored with this noninvasive, nonionizing technology, such as hyperthermia generation, to release drugs from thermosensitive liposomal carriers. These drugs can include chemotherapies such as doxorubicin, for which targeted release is desired due to the dose-limiting systemic side effects, namely cardiotoxicity. Doxorubicin is a mainstay for treating a variety of malignant tumors and is commonly used in relapsed or recurrent rhabdomyosarcoma (RMS). RMS is the most common solid soft tissue extracranial tumor in children and young adults. Despite aggressive, multimodal therapy, RMS survival rates have remained the same for the past 30 years. To explore a solution for addressing this unmet need, an experimental protocol was developed to evaluate the release of thermosensitive liposomal doxorubicin (TLD) in an immunocompetent, syngeneic RMS mouse model using MRgHIFU as the source of hyperthermia for drug release.

Presented here is a protocol to use controlled hyperthermia, generated by magnetic resonance-guided high intensity focused ultrasound, to trigger drug release from temperature-sensitive liposomes in a rhabdomyosarcoma mouse model.

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time ...