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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Cancer Research

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

Published: March 24th, 2022

DOI:

10.3791/62560

1Department of Diagnostic Sciences, Ghent University, 2Department of Electronics and Information Systems, Ghent University, 3Department of Head and Skin, Ghent University

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 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.

Glioblastoma (GB) is a malignant and very aggressive primary brain tumor. GB is a solid heterogeneous tumor typically characterized by infiltrative boundaries, nuclear atypia, and necrosis1. The presence of the blood-brain-barrier and the brain's status as an immune-privileged site makes the discovery of novel targets for chemo- and immunotherapy a daunting task2,3,4. It is noteworthy that the treatment of GB patients has barely changed since the introduction, in 2005, of the Stupp protocol that combines external beam radiation therapy (RT) with co....

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The study was approved by the local ethics committee for animal experiments (ECD 18/21). Anesthesia monitoring is performed by acquiring the respiratory rate of the animals using a sensor.

1. F98 GB rat cell model

  1. Culture the F98 GB cells in a monolayer using Dulbecco's Modified Eagle Medium, supplemented with 10% calf serum, 1% penicillin, 1% streptomycin, and 1% L-glutamine, and place them in a CO2 incubator (5% CO2 and 37 °C).
  2. Inocul.......

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The feasibility of PET- and MRI-guided irradiation in a glioblastoma rat model using the SARRP to mimic the human treatment strategy has been previously described20,21,22. While the animal was fixed on a multimodality bed made in-house, it was possible to create an acceptable radiation treatment plan combining three imaging modalities: PET, MRI, and CT. In these methods, an external software package (see the Table of Mat.......

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A rat GB model to mimic the chemo-radiation treatment in the clinic for glioblastoma patients was previously described20. Similar to the clinical method, CT and MRI were combined during the treatment-planning process to obtain more precise irradiation. A multimodality bed to minimize (head) movement was used when the animal was moved from one imaging system to another. Subsequently, PET imaging was added to the treatment-planning process, and PET-based sub-volume boosting could be successfully imp.......

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The authors would like to thank Lux Luka Foundation for supporting this work.

....

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Name Company Catalog Number Comments
Cell culture
F98 Glioblastoma Cell Line ATCC CRL-2397 https://www.lgcstandards-atcc.org/products/all/CRL-2397
Dulbeco's Modified Eagle Medium Thermo Fisher Scientific 22320-030
Cell culture flasks Thermo Fisher Scientific 178883 75 cm²
FBS Thermo Fisher Scientific 10270106
L-Glutamine Thermo Fisher Scientific 25030-032 200 mM
Penicilline-Streptomycin Thermo Fisher Scientific 15140-148 10,000 U/mL
Phosphate-Buffered Saline (PBS) Thermo Fisher Scientific 14040-224
Trypsin-EDTA Thermo Fisher Scientific 25300-062 0.05%
GB Rat Model
Ball-shaped burr Foredom A-228 1.8 mm
Bone Wax Aesculap 1029754 https://www.aesculapusa.com/en/healthcare-professionals/or-solutions/or-solutions-cranial-closure/hemostatic-bone-wax.html
Ethilon Ethicon 662G/662H FS-2, 4-0, 3/8, 19 mm
Fischer F344/Ico crl Rats Charles River -
Insulin Syringe Microfine Beckton-Dickinson 320924 1 mL, 29 G
IR Lamp Philips HP3616/01
Meloxicam (Metacam) Boehringer Ingelheim - 2 mg/mL
Micromotor rotary tool Foredom K.1090-22
Micropump system Stoelting Co. 53312 Stoelting Stereotaxic Injector
Stereotactic frame Stoelting Co. 51600
Xylocaine (1%, with adrenaline 1:200,000) Aspen - 1%, with adrenaline 1:200,000
Xylocaine gel (2%) Aspen - 2%
Animal Irradiation
Micro-irradiator X-Strahl SARRP Version 4.2.0
Software X-Strahl Muriplan Preclinical treatment planning system (PCTPC), version 2.2.2
Small Animal PET
[18F]FET Inhouse made - PET tracer; along with Prohance: MRI/PET agent
Micro-PET Molecubes Beta-Cube https://www.molecubes.com/b-cube/
Small Animal MRI
Micro-MRI Bruker Biospin Pharmascan 70/16 https://www.bruker.com/products/mr/preclinical-mri/pharmascan.html
30 G Needle for IV injection Beckton-Dickinson 305128
PE 10 Tubing Instech Laboratories Inc BTPE-10 BTPE-10, polyethylene tubing 0.011 x 0.024 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
Prohance contrast agent Bracco Imaging - 279.3 mg/mL, gadolinium-contrast agent (along with [18F]FET: MRI/PET agent)
Tx/Rx Rat Brain - Mouse Whole Body Volumecoil Bruker Biospin - 40 mm diameter
Water-based Heating Unit Bruker Biospin MT0125
Consumables
Isoflurane Zoetis B506 Anesthesia
Insulin Syringe Microfine Beckton-Dickinson 320924 1 mL, 29 G
Image Analysis
MATLAB Mathworks - Version R2019b
PMOD PMOD technologies LLC Preclinical and molecular imaging software

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