Stability analysis of patient-specific 4DCT- and 4DCBCT-based correspondence models.

BACKGROUND: Surrogate-based motion compensation in stereotactic body radiation therapy (SBRT) strongly relies on a constant relationship between an external breathing signal and the internal tumor motion over the course of treatment, that is, a stable patient-specific correspondence model. PURPOSE: This study aims to develop methods for analyzing the stability of correspondence models by integrating planning 4DCT and pretreatment 4D cone-beam computed tomography (4DCBCT) data and assessing the relation to patient-specific clinical parameters. METHODS: For correspondence modeling, a regression-based approach is applied, correlating patient-specific internal motion (vector fields computed by deformable image registration) and external breathing signals (recorded by Varian's RPM and RGSC system). To analyze correspondence model stability, two complementary methods are proposed. (1) Target volume-based analysis: 4DCBCT-based correspondence models predict clinical target volumes (GTV and internal target volume [ITV]) within the planning 4DCT, which are evaluated by overlap and distance measures (Dice similarity coefficient [DSC]/average symmetric surface distance [ASSD]). (2) System matrix-based analysis: 4DCBCT-based regression models are compared to 4DCT-based models using mean squared difference (MSD) and principal component analysis of the system matrices. Stability analysis results are correlated with clinical parameters. Both methods are applied to a dataset of 214 pretreatment 4DCBCT scans (Varian TrueBeam) from a cohort of 46 lung tumor patients treated with ITV-based SBRT (planning 4DCTs acquired with Siemens AS Open and SOMATOM go.OPEN Pro CT scanners). RESULTS: Consistent results across the two complementary analysis approaches (Spearman correlation coefficient of 0.6 / 0.7 $0.6/ 0.7$ between system matrix-based MSD and GTV-based DSC/ASSD) were observed. Analysis showed that stability was not predominant, with 114/214 fraction-wise models not surpassing a threshold of D S C > 0.7 $DSC > 0.7$ for the GTV, and only 14/46 patients demonstrating a D S C > 0.7 $DSC > 0.7$ in all fractions. Model stability did not degrade over the course of treatment. The mean GTV-based DSC is 0.59 ± 0.26 $0.59\pm 0.26$ (mean ASSD of 2.83 ± 3.37 $2.83\pm 3.37$ ) and the respective ITV-based DSC is 0.69 ± 0.20 $0.69\pm 0.20$ (mean ASSD of 2.35 ± 1.81 $2.35\pm 1.81$ ). The clinical parameters showed a strong correlation between smaller tumor motion ranges and increased stability. CONCLUSIONS: The proposed methods identify patients with unstable correspondence models prior to each treatment fraction, serving as direct indicators for the necessity of replanning and adaptive treatment approaches to account for internal-external motion variations throughout the course of treatment.

Time-resolved role of P2X4 and P2X7 during CD8+ T cell activation.

CD8+ T cells are a crucial part of the adaptive immune system, responsible for combating intracellular pathogens and tumor cells. The initial activation of T cells involves the formation of highly dynamic Ca2+ microdomains. Recently, purinergic signaling was shown to be involved in the formation of the initial Ca2+ microdomains in CD4+ T cells. In this study, the role of purinergic cation channels, particularly P2X4 and P2X7, in CD8+ T cell signaling from initial events to downstream responses was investigated, focusing on various aspects of T cell activation, including Ca2+ microdomains, global Ca2+ responses, NFAT-1 translocation, cytokine expression, and proliferation. While Ca2+ microdomain formation was significantly reduced in the first milliseconds to seconds in CD8+ T cells lacking P2X4 and P2X7 channels, global Ca2+ responses over minutes were comparable between wild-type (WT) and knockout cells. However, the onset velocity was reduced in P2X4-deficient cells, and P2X4, as well as P2X7-deficient cells, exhibited a delayed response to reach a certain level of free cytosolic Ca2+ concentration ([Ca2+]i). NFAT-1 translocation, a crucial transcription factor in T cell activation, was also impaired in CD8+ T cells lacking P2X4 and P2X7. In addition, the expression of IFN-γ, a major pro-inflammatory cytokine produced by activated CD8+ T cells, and Nur77, a negative regulator of T cell activation, was significantly reduced 18h post-stimulation in the knockout cells. In line, the proliferation of T cells after 3 days was also impaired in the absence of P2X4 and P2X7 channels. In summary, the study demonstrates that purinergic signaling through P2X4 and P2X7 enhances initial Ca2+ events during CD8+ T cell activation and plays a crucial role in regulating downstream responses, including NFAT-1 translocation, cytokine expression, and proliferation on multiple timescales. These findings suggest that targeting purinergic signaling pathways may offer potential therapeutic interventions.

Benchmarking machine learning-based real-time respiratory signal predictors in 4D SBRT.

BACKGROUND: Stereotactic body radiotherapy of thoracic and abdominal tumors has to account for respiratory intrafractional tumor motion. Commonly, an external breathing signal is continuously acquired that serves as a surrogate of the tumor motion and forms the basis of strategies like breathing-guided imaging and gated dose delivery. However, due to inherent system latencies, there exists a temporal lag between the acquired respiratory signal and the system response. Respiratory signal prediction models aim to compensate for the time delays and to improve imaging and dose delivery. PURPOSE: The present study explores and compares six state-of-the-art machine and deep learning-based prediction models, focusing on real-time and real-world applicability. All models and data are provided as open source and data to ensure reproducibility of the results and foster reuse. METHODS: The study was based on 2502 breathing signals ( t t o t a l ≈ 90 $t_{total} \approx 90$  h) acquired during clinical routine, split into independent training (50%), validation (20%), and test sets (30%). Input signal values were sampled from noisy signals, and the target signal values were selected from corresponding denoised signals. A standard linear prediction model (Linear), two state-of-the-art models in general univariate signal prediction (Dlinear, Xgboost), and three deep learning models (Lstm, Trans-Enc, Trans-TSF) were chosen. The prediction performance was evaluated for three different prediction horizons (480, 680, and 920 ms). Moreover, the robustness of the different models when applied to atypical, that is, out-of-distribution (OOD) signals, was analyzed. RESULTS: The Lstm model achieved the lowest normalized root mean square error for all prediction horizons. The prediction errors only slightly increased for longer horizons. However, a substantial spread of the error values across the test signals was observed. Compared to typical, that is, in-distribution test signals, the prediction accuracy of all models decreased when applied to OOD signals. The more complex deep learning models Lstm and Trans-Enc showed the least performance loss, while the performance of simpler models like Linear dropped the most. Except for Trans-Enc, inference times for the different models allowed for real-time application. CONCLUSION: The application of the Lstm model achieved the lowest prediction errors. Simpler prediction filters suffer from limited signal history access, resulting in a drop in performance for OOD signals.