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Breast-conserving surgery requires supportive radiotherapy to prevent cancer recurrence. However, the task of localizing the tumor bed to be irradiated is not trivial. The automatic image registration could significantly aid the tumor bed localization and lower the radiation dose delivered to the surrounding healthy tissues. This study proposes a novel image registration method dedicated to breast tumor bed localization addressing the problem of missing data due to tumor resection that may be applied to real-time radiotherapy planning. We propose a deep learning-based nonrigid image registration method based on a modified U-Net architecture. The algorithm works simultaneously on several image resolutions to handle large deformations. Moreover, we propose a dedicated volume penalty that introduces the medical knowledge about tumor resection into the registration process. The proposed method may be useful for improving real-time radiation therapy planning after the tumor resection and, thus, lower the surrounding healthy tissues’ irradiation. The data used in this study consist of 30 computed tomography scans acquired in patients with diagnosed breast cancer, before and after tumor surgery. The method is evaluated using the target registration error between manually annotated landmarks, the ratio of tumor volume, and the subjective visual assessment. We compare the proposed method to several other approaches and show that both the multilevel approach and the volume regularization improve the registration results. The mean target registration error is below 6.5 mm, and the relative volume ratio is close to zero. The registration …