Abstract
Background
Radiotherapy induces systemic changes beyond the targeted tumour site, yet the biological mechanisms driving these effects remain poorly understood. This study aims to longitudinally profile acute systemic plasma proteomic responses to radiotherapy in patients with prostate, bladder, and head and neck cancers, providing insight into shared and tumour-specific effects and identifying biomarkers predictive of treatment-related toxicities.
Methods
We apply our previously developed Nano-proteomics workflow to comprehensively analyse longitudinal weekly plasma samples collected before and during radiotherapy. We perform mass spectrometry-based differential protein analysis to identify acute changes of the plasma proteome and enriched biological pathways involved. In the prostate cohort, we associate plasma proteomics with clinical toxicity outcomes.
Results
Our data indicate that the most significant systemic proteomic changes occur within the first two weeks of radiotherapy, highlighting a critical period for biomarker identification. Across all patient cohorts, we observe common biological responses: rapid activation of inflammatory and immune pathways, followed by structural reorganisation and immune resolution, regardless of tumour type or concurrent treatments. Despite this shared acute response, distinct protein mediators are found to be dysregulated in a tumour-specific manner. In the prostate cancer cohort, plasma profiling at baseline, one week after the initiation of radiotherapy, and at the end of radiotherapy, results in the identification of 28, 29, and 20 proteins, respectively, that are associated with subsequent bowel and urinary toxicities.
Conclusions
This study underscores the value of longitudinal proteomics in uncovering systemic effects of radiotherapy and supports the potential of plasma proteomic biomarkers to identify patients at increased risk of radiotherapy-induced toxicity, paving the way for personalised treatment strategies.
Plain language summary
Radiotherapy targets tumour cells but also triggers systemic effects beyond the irradiated site. To characterise these responses, we analysed blood samples from patients with prostate, bladder, and head and neck cancers collected before and during treatment. Using a nanoparticle-enabled proteomics workflow, we profiled weekly changes in circulating proteins and observed the strongest systemic alterations within the first two weeks of radiotherapy. While many systemic responses were shared across cancer types, tumour-specific patterns also emerged. In the prostate cancer cohort, longitudinal plasma profiles revealed proteins linked to later bowel and urinary toxicities, suggesting potential predictive biomarkers. Overall, our findings underscore the systemic nature of radiotherapy and highlight longitudinal plasma proteomics as a valuable tool for biomarker discovery and personalised treatment.
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Data availability
The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [1] partner repository with the dataset identifier PXD071492 and 10.6019/PXD071492. The source data for Fig. 3(a-b), Fig. 4g–I and for the supporting Figures S1b, S2b, S3a–c, and S5a are provided in Supplementary Data set 2.
Code availability
The underlying code for this study is available at https://github.com/assisalam/nanoOmic/ [github.com]. Computational method summary table is provided in Supplementary Data 5.
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Acknowledgements
The clinical research was carried out at the National Institute for Health and Care Research (NIHR) Manchester Biomedical Research Centre (BRC) (NIHR203308). The proteomic analyses and DCW were supported by Cancer Research UK RadNet Manchester [C1994/A28701]. We thank the Faculty of Biology, Medicine and Health, University of Manchester Biological Mass Spectrometry Facility (Bio-MS) staff for their support. We thank the Manchester Cancer Research Centre biobank (MCRC biobank). E.C., CGQ & DCW are funded by the NIHR Biomedical Research Centre (NIHR203308). A.C. and C.W. are supported by the NIHR Manchester Biomedical Research Centre. We would like to thank the research nurse team at the Christie Hospital.
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H.Ab. initiated and performed the proteomic experiments, analysed the data, and wrote the manuscript. S.A., X.L., and C.G. contributed to the proteomics data analysis. E.Ch. and T.L. provided clinical samples and clinical information. C.W. designed and was Chief Investigator of the clinical study. A.C. was involved in patient recruitment. K.J.W., E.C., K.K., A.C., D.W., and C.W. supervised the work and contributed to the design of the study, interpretation of the data and the revising of the manuscript. M.H. contributed to the conceptualization of the study, designed the experiments, supervised the execution of the proteomics experimental work and data analysis, contributed to the interpretation of the data, and prepared and reviewed the manuscript.
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Abumanhal-Masarweh, H., Assi, S.A., Liu, X. et al. Longitudinal plasma nano-proteomics reveals acute systemic responses to radiotherapy and predictive biomarkers of late toxicity. Commun Med (2026). https://doi.org/10.1038/s43856-026-01552-3
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DOI: https://doi.org/10.1038/s43856-026-01552-3
