A precision 3D conformal treatment technique in rats: Application to whole-brain radiotherapy with hippocampal avoidance.

PURPOSE: To develop and validate three-dimensional (3D) conformal hippocampal sparing whole-brain radiation therapy (HA-WBRT) for Wistar rats utilizing precision 3D-printed immobilization and micro-blocks. This technique paves the way for future preclinical studies investigating brain treatments that reduce neurotoxicity. METHODS AND MATERIALS: A novel preclinical treatment planning and delivery process was developed to enable precision 3D conformal treatment and hippocampal avoidance capability for the Xrad 225cx small animal irradiator. A range of conformal avoidance plans were evaluated consisting of equiangularly spaced coplanar axial beams, with plans containing 2, 4, 7, and 8 fields. The hippocampal sparing and coverage of these plans were investigated through Monte Carlo dose calculation (SmART-Plan Xrad 225cx planning system). Treatment delivery was implemented through a novel process where hippocampal block shapes were computer generated from an MRI rat atlas which was registered to on-board cone beam CT of the rat in treatment position. The blocks were 3D printed with a tungsten-doped filament at lateral resolution of 80 µm. Precision immobilization was achieved utilizing a 3D-printed support system which enabled angled positioning of the rat head in supine position and bite block to improve coverage of the central diencephalon. Treatment delivery was verified on rodent-morphic Presage® 3D dosimeters optically scanned at 0.2-mm isotropic resolution. Biological verification of hippocampal avoidance was performed with immunohistologic staining. RESULTS: All simulated plans spared the hippocampus while delivering high dose to the brain (22.5-26.2 Gy mean dose to brain at mean hippocampal dose of 7 Gy). No significant improvement in hippocampal sparing was observed by adding beams beyond four fields. Dosimetric sparing of hippocampal region of the four-field plan was verified with the Presage® dosimeter (mean dose = 9.6 Gy, D100% = 7.1 Gy). Simulation and dosimeter match at distance-to-agreement of 2 mm and dose difference of ±3% at 91.7% gamma passing rate (passing criteria of γ < 1). Agreement is less at 1 mm and ±5% at 69.0% gamma passing rate. The four-field plan was further validated with immunohistochemistry and showed a significant reduction in DNA double-strand breaks within the spared region compared with whole-brain irradiated groups (P = 0.021). However, coverage of the whole brain was low at 48.5-57.8% of the volume receiving 30Gy at 7Gy mean hippocampal dose in simulation and 46.7-52.5% in dosimetric measurements. This can be attributed to the shape of the rat hippocampus and the inability of treatment platform to employ non-coplanar beams. CONCLUSION: A novel approach for conformal microradiation therapy using 3D-printing technology was developed, implemented, and validated. A workflow was developed to generate accurate 3D-printed blocks from registered high-resolution rat MRI atlas structures. Although hippocampus was spared with this technique, whole-brain target coverage was suboptimal, indicating that non-coplanar beams and IMRT capability may be required to meet stringent dose criteria associated with current human RTOG trials.