BACKGROUND: Management of high-grade gliomas (HGGs) close to motor areas is challenging due to the risk of treatment-related morbidity. Thus, for resection, functional mapping of the corticospinal tract (CST) with navigated transcranial magnetic stimulation (nTMS) combined with diffusion tensor imaging (DTI)-based fiber tracking (nTMS-DTI-FT) is increasingly used. This study investigated the application of nTMS-DTI-FT in adjuvant radiation therapy (RT) planning of HGGs for CST avoidance. METHODS: The preoperative nTMS-DTI-FT based CST of 35 patients harboring HGGs were incorporated into the RT planning system and merged with planning imaging. The CST was delineated as PRV-FTTMS. Intensity-modulated RT (IMRT) plans were optimized to preserve PRV-FTTMS. Segments within the planning target volume (PTV) were not spared (overlap). RESULTS: With plan optimization, mean dose (Dmean) of PRV-FTTMS can be reduced by 17.1% on average (range 0.1 - 37.9%), thus from 25.5 Gy to 21.2 Gy (p<0.001 For PRV-FTTMS segments beyond the PTV dose, reduction is possible by 26.8% (range 0.1 - 43.9%, Dmean 17.4 Gy vs. 12.5 Gy, p<0.001 Considering only portions within the 50% isodose level, Dmean is decreased by 46.7% from 38.6 Gy to 20.5 Gy (range 19.1 - 62.8%, p<0.001. PTV coverage was not affected: V95% and V90% were 96.4 ± 3.1% and 98.0 ± 3.9% vs. 96.1 ± 3.5% (p=0.34) and 98.3 ± 2.9% (p=0.58). Dose constraints for organs at risk (OARs) were all met. CONCLUSION: This study demonstrates that nTMS-DTI-FT can be utilized in the RT planning of HGGs for CST sparing. However, the degree of dose reduction depends on the overlap with the PTV. The functional benefit needs to be investigated in future prospective clinical trials.