{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T13:51:00Z","timestamp":1768830660198,"version":"3.49.0"},"reference-count":41,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2025,6,26]],"date-time":"2025-06-26T00:00:00Z","timestamp":1750896000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia (FCT)","award":["PTDC\/MED-OUT\/31118\/2017"],"award-info":[{"award-number":["PTDC\/MED-OUT\/31118\/2017"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia (FCT)","award":["CEEC-INST\/00106\/2021"],"award-info":[{"award-number":["CEEC-INST\/00106\/2021"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia (FCT)","award":["UID\/00306\/2025"],"award-info":[{"award-number":["UID\/00306\/2025"]}]},{"name":"FCT","award":["PTDC\/MED-OUT\/31118\/2017"],"award-info":[{"award-number":["PTDC\/MED-OUT\/31118\/2017"]}]},{"name":"FCT","award":["CEEC-INST\/00106\/2021"],"award-info":[{"award-number":["CEEC-INST\/00106\/2021"]}]},{"name":"FCT","award":["UID\/00306\/2025"],"award-info":[{"award-number":["UID\/00306\/2025"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pre-metastatic niche (PMN) formation is a critical step in metastatic progression. However, the biological effects of subtherapeutic doses of ionizing radiation (SDIRs) following radiotherapy on this process remain unclear. Using a 4T1 breast cancer mouse model, we investigated the effects of SDIRs (3 \u00d7 0.3 Gy) on lung PMN development and metastasis upon SDIR exposure on days 8\u201310 post-tumor injection, followed by mastectomy and analyzed on day 24. SDIRs significantly increased the total metastatic volume (TMV) in lungs, suggesting an accelerated PMN formation. Mechanistically, the SDIR acted as an early catalyst for niche priming, upregulating Bv8 expression, enhancing neutrophil recruitment, and increasing MMP9, S100A8, and Il6 production in the PMN by day 11. Moreover, SDIR drives metastasis through distinct mechanisms. Proteomic analysis revealed SDIR-driven metabolic reprogramming, with a shift away from fatty acid metabolism toward glycolysis and lipid accumulation within the PMN. This shift contributes to extracellular matrix (ECM) remodeling, immune modulation, and the upregulation of adhesion-related pathways, shaping a microenvironment that accelerates metastatic outgrowth. By reprogramming the pre-metastatic lung, the SDIR highlights the need to integrate organ-specific radiation exposure into metastasis models. Metabolic and immune-stromal pathways emerge as potential therapeutic targets, underscoring the importance of refining radiotherapy strategies to mitigate unintended pro-metastatic effects.<\/jats:p>","DOI":"10.3390\/ijms26136145","type":"journal-article","created":{"date-parts":[[2025,6,26]],"date-time":"2025-06-26T06:55:13Z","timestamp":1750920913000},"page":"6145","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Subtherapeutic Dose of Ionizing Radiation Reprograms the Pre-Metastatic Lung Niche, Accelerating Its Formation and Promoting Metastasis"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1721-9909","authenticated-orcid":false,"given":"Paula de","family":"Oliveira","sequence":"first","affiliation":[{"name":"Centro Cardiovascular da Universidade de Lisboa (CCUL@RISE), Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal"}]},{"given":"In\u00eas Sofia","family":"Vala","sequence":"additional","affiliation":[{"name":"Centro Cardiovascular da Universidade de Lisboa (CCUL@RISE), Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3922-3602","authenticated-orcid":false,"given":"Pedro","family":"Fa\u00edsca","sequence":"additional","affiliation":[{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1664-472X","authenticated-orcid":false,"given":"Joao C","family":"Guimaraes","sequence":"additional","affiliation":[{"name":"Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal"}]},{"given":"Filomena","family":"Pina","sequence":"additional","affiliation":[{"name":"Santa Maria Hospital, Unidade Local de Sa\u00fade de Santa Maria (ULSSM), 1649-035 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7544-7726","authenticated-orcid":false,"given":"Esmeralda","family":"Poli","sequence":"additional","affiliation":[{"name":"Santa Maria Hospital, Unidade Local de Sa\u00fade de Santa Maria (ULSSM), 1649-035 Lisboa, Portugal"}]},{"given":"Isabel","family":"Diegues","sequence":"additional","affiliation":[{"name":"Santa Maria Hospital, Unidade Local de Sa\u00fade de Santa Maria (ULSSM), 1649-035 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6362-8255","authenticated-orcid":false,"given":"Hugo","family":"Os\u00f3rio","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4099-002 Porto, Portugal"},{"name":"IPATIMUP\u2014Instituto de Patologia e Imunologia Molecular, Universidade do Porto, 4099-002 Porto, Portugal"},{"name":"FMUP\u2014Faculdade de Medicina, Universidade do Porto, 4099-002 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6353-2616","authenticated-orcid":false,"given":"Rune","family":"Matthiesen","sequence":"additional","affiliation":[{"name":"Computational and Experimental Biology Group, iNOVA4Health, NOVA Medical School, Faculdade de Ci\u00eancias M\u00e9dicas, Universidade NOVA de Lisboa, 1349-008 Lisboa, Portugal"}]},{"given":"Karine","family":"Serre","sequence":"additional","affiliation":[{"name":"Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5711-1292","authenticated-orcid":false,"given":"Susana","family":"Constantino Rosa Santos","sequence":"additional","affiliation":[{"name":"Centro Cardiovascular da Universidade de Lisboa (CCUL@RISE), Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1144","DOI":"10.1016\/S1470-2045(15)00285-5","article-title":"Global Task Force on Radiotherapy for Cancer Control","volume":"16","author":"Jaffray","year":"2015","journal-title":"Lancet Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"309","DOI":"10.3322\/caac.21341","article-title":"Cardiotoxicity of anticancer treatments: Epidemiology, detection, and management","volume":"66","author":"Curigliano","year":"2016","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1007\/s00066-022-02032-y","article-title":"Systematic risk analysis of radiation pneumonitis in breast cancer: Role of cotreatment with chemo-, endocrine, and targeted therapy","volume":"199","author":"Mangesius","year":"2023","journal-title":"Strahlenther. Onkol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ribeiro, S., Simoes, A.R., Rocha, F., Vala, I.S., Pinto, A.T., Ministro, A., Poli, E., Diegues, I.M., Pina, F., and Benadjaoud, M.A. (2022). Molecular Changes In Cardiac Tissue As A New Marker To Predict Cardiac Dysfunction Induced By Radiotherapy. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.945521"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.radonc.2020.06.038","article-title":"Low doses of ionizing radiation activate endothelial cells and induce angiogenesis in peritumoral tissues","volume":"151","author":"Poli","year":"2020","journal-title":"Radiother. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Marques, F.G., Carvalho, L., Sousa, J.S., Rino, J., Diegues, I., Poli, E., Pina, F., Saude, L., and Constantino Rosa Santos, S. (2020). Low doses of ionizing radiation enhance angiogenesis and consequently accelerate post-embryonic development but not regeneration in zebrafish. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-60129-9"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1667\/RR15438.1","article-title":"Low Doses of Ionizing Radiation Enhance the Angiogenic Potential of Adipocyte Conditioned Medium","volume":"192","author":"Marques","year":"2019","journal-title":"Radiat. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1093\/cvr\/cvx065","article-title":"Low-dose ionizing radiation induces therapeutic neovascularization in a pre-clinical model of hindlimb ischemia","volume":"113","author":"Ministro","year":"2017","journal-title":"Cardiovasc. Res."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Sofia Vala, I., Martins, L.R., Imaizumi, N., Nunes, R.J., Rino, J., Kuonen, F., Carvalho, L.M., Ruegg, C., Grillo, I.M., and Barata, J.T. (2010). Low doses of ionizing radiation promote tumor growth and metastasis by enhancing angiogenesis. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0011222"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"26615","DOI":"10.18632\/oncotarget.5666","article-title":"Radiation-induced lung damage promotes breast cancer lung-metastasis through CXCR4 signaling","volume":"6","author":"Feys","year":"2015","journal-title":"Oncotarget"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2040","DOI":"10.1016\/j.cmet.2021.09.002","article-title":"Tumor-derived exosomes drive immunosuppressive macrophages in a pre-metastatic niche through glycolytic dominant metabolic reprogramming","volume":"33","author":"Morrissey","year":"2021","journal-title":"Cell Metab."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1038\/s41590-018-0229-5","article-title":"Unique pattern of neutrophil migration and function during tumor progression","volume":"19","author":"Patel","year":"2018","journal-title":"Nat. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1038\/nrc.2017.6","article-title":"Pre-metastatic niches: Organ-specific homes for metastases","volume":"17","author":"Peinado","year":"2017","journal-title":"Nat. Rev. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Chen, E., and Yu, J. (2023). The role and metabolic adaptations of neutrophils in premetastatic niches. Biomark. Res., 11.","DOI":"10.1186\/s40364-023-00493-6"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5045","DOI":"10.1038\/onc.2017.118","article-title":"CYP4A in tumor-associated macrophages promotes pre-metastatic niche formation and metastasis","volume":"36","author":"Chen","year":"2017","journal-title":"Oncogene"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Doglioni, G., Parik, S., and Fendt, S.M. (2019). Interactions in the (Pre)metastatic Niche Support Metastasis Formation. Front. Oncol., 9.","DOI":"10.3389\/fonc.2019.00219"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1900195","DOI":"10.1002\/advs.201900195","article-title":"Macrophages-Triggered Sequential Remodeling of Endothelium-Interstitial Matrix to Form Pre-Metastatic Niche in Microfluidic Tumor Microenvironment","volume":"6","author":"Kim","year":"2019","journal-title":"Adv. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1038\/s41568-024-00752-0","article-title":"Cancer-induced systemic pre-conditioning of distant organs: Building a niche for metastatic cells","volume":"24","author":"Rabas","year":"2024","journal-title":"Nat. Rev. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2162","DOI":"10.1038\/s41388-018-0564-x","article-title":"Macrophage-mediated transfer of cancer-derived components to stromal cells contributes to establishment of a pro-tumor microenvironment","volume":"38","author":"Umakoshi","year":"2019","journal-title":"Oncogene"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Wang, Y., Ding, Y., Guo, N., and Wang, S. (2019). MDSCs: Key Criminals of Tumor Pre-metastatic Niche Formation. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.00172"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1186\/s41232-022-00208-8","article-title":"Inflammation-associated premetastatic niche formation","volume":"42","author":"Deguchi","year":"2022","journal-title":"Inflamm. Regen."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1002\/ijc.29444","article-title":"The lack of type I interferon induces neutrophil-mediated pre-metastatic niche formation in the mouse lung","volume":"137","author":"Wu","year":"2015","journal-title":"Int. J. Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Wu, M., Ma, M., Tan, Z., Zheng, H., and Liu, X. (2020). Neutrophil: A New Player in Metastatic Cancers. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.565165"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.jconrel.2020.06.037","article-title":"Prognosis and targeting of pre-metastatic niche","volume":"325","author":"Zhou","year":"2020","journal-title":"J. Control. Release"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1038\/s41577-018-0066-7","article-title":"Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer","volume":"18","author":"Jones","year":"2018","journal-title":"Nat. Rev. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Li, R., Wen, A., and Lin, J. (2020). Pro-Inflammatory Cytokines in the Formation of the Pre-Metastatic Niche. Cancers, 12.","DOI":"10.3390\/cancers12123752"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4977","DOI":"10.1038\/s41467-020-18701-4","article-title":"Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during clinical latency","volume":"11","author":"Grujovic","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1186\/s12943-024-02107-7","article-title":"Neutrophils in the premetastatic niche: Key functions and therapeutic directions","volume":"23","author":"Jia","year":"2024","journal-title":"Mol. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1038\/s41392-024-01937-7","article-title":"Pre-metastatic niche: Formation, characteristics and therapeutic implication","volume":"9","author":"Wang","year":"2024","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.tem.2024.07.016","article-title":"Adapt and shape: Metabolic features within the metastatic niche","volume":"36","author":"Pranzini","year":"2025","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.cmet.2013.05.017","article-title":"Cellular fatty acid metabolism and cancer","volume":"18","author":"Currie","year":"2013","journal-title":"Cell Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1038\/s41467-019-13668-3","article-title":"Mitochondrial TCA cycle metabolites control physiology and disease","volume":"11","author":"Chandel","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Nolfi-Donegan, D., Braganza, A., and Shiva, S. (2020). Mitochondrial electron transport chain: Oxidative phosphorylation, oxidant production, and methods of measurement. Redox Biol., 37.","DOI":"10.1016\/j.redox.2020.101674"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.1038\/ncb1507","article-title":"Tumour-mediated upregulation of chemoattractants and recruitment of myeloid cells predetermines lung metastasis","volume":"8","author":"Hiratsuka","year":"2006","journal-title":"Nat. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1007\/s00018-015-2070-4","article-title":"Reprogramming of glucose, fatty acid and amino acid metabolism for cancer progression","volume":"73","author":"Li","year":"2016","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1038\/s41590-020-0783-5","article-title":"Lung mesenchymal cells elicit lipid storage in neutrophils that fuel breast cancer lung metastasis","volume":"21","author":"Li","year":"2020","journal-title":"Nat. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/s12943-025-02258-1","article-title":"Lipid metabolic reprograming: The unsung hero in breast cancer progression and tumor microenvironment","volume":"24","author":"Wan","year":"2025","journal-title":"Mol. Cancer"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1016\/j.ccell.2023.02.018","article-title":"Immune determinants of the pre-metastatic niche","volume":"41","author":"Patras","year":"2023","journal-title":"Cancer Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/s41596-018-0082-x","article-title":"Single-pot, solid-phase-enhanced sample preparation for proteomics experiments","volume":"14","author":"Hughes","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3294","DOI":"10.1074\/mcp.M113.034363","article-title":"Global mass spectrometry and transcriptomics array based drug profiling provides novel insight into glucosamine induced endoplasmic reticulum stress","volume":"13","author":"Carvalho","year":"2014","journal-title":"Mol. Cell. Proteom."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"108967","DOI":"10.1016\/j.celrep.2021.108967","article-title":"Self-renewal of double-negative 3 early thymocytes enables thymus autonomy but compromises the beta-selection checkpoint","volume":"35","author":"Paiva","year":"2021","journal-title":"Cell Rep."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6145\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:59:05Z","timestamp":1760032745000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6145"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,26]]},"references-count":41,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["ijms26136145"],"URL":"https:\/\/doi.org\/10.3390\/ijms26136145","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,26]]}}}