{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T12:54:05Z","timestamp":1791377645250,"version":"4.3.2"},"reference-count":31,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,12]],"date-time":"2025-05-12T00:00:00Z","timestamp":1747008000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Mary Kay Ash Foundation","award":["CA014089"],"award-info":[{"award-number":["CA014089"]}]},{"name":"Mary Kay Ash Foundation","award":["CA023074"],"award-info":[{"award-number":["CA023074"]}]},{"name":"Cleveland Clinic and by ANGLE plc (Stage IV cohort)","award":["CA014089"],"award-info":[{"award-number":["CA014089"]}]},{"name":"Cleveland Clinic and by ANGLE plc (Stage IV cohort)","award":["CA023074"],"award-info":[{"award-number":["CA023074"]}]},{"name":"Society of Surgical Oncology Clinical Investigator Award","award":["CA014089"],"award-info":[{"award-number":["CA014089"]}]},{"name":"Society of Surgical Oncology Clinical Investigator Award","award":["CA023074"],"award-info":[{"award-number":["CA023074"]}]},{"name":"a STOP Cancer Marni Levine Memorial Seed Grant (JL) for the early-stage breast cancer cohort","award":["CA014089"],"award-info":[{"award-number":["CA014089"]}]},{"name":"a STOP Cancer Marni Levine Memorial Seed Grant (JL) for the early-stage breast cancer cohort","award":["CA023074"],"award-info":[{"award-number":["CA023074"]}]},{"name":"National Cancer Institute","award":["CA014089"],"award-info":[{"award-number":["CA014089"]}]},{"name":"National Cancer Institute","award":["CA023074"],"award-info":[{"award-number":["CA023074"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background: The mechanical conditioning (MeCo) score is a multigene expression signature that is acquired by cancer cells in the primary breast tumor and is reflective of their responsiveness to ECM stiffness caused by tumor fibrosis. Chromatin remodeling downstream of mechanotransduction allows cancer cells to retain these acquired aggressive features even in the absence of mechanical stimulation from the primary tumor microenvironment, for instance, after dissemination through systemic circulation during metastasis. Importantly, patients who have high MeCo score tumors are at higher risk of developing metastatic breast cancer, compared to those with low MeCo scores. Moreover, circulating tumor cells (CTCs) are associated with a higher rate of metastatic dissemination, making CTC detection in the circulation of patients with breast cancer a significant prognostic biomarker for breast cancer metastasis. Beyond their enumeration per blood volume units, specific prognostic features of CTCs are not fully explored. We sought to determine whether MeCo scores increase stepwise along the metastatic cascade, from primary tumors to CTCs to distant metastatic colonization, using patient-matched biopsies. Methods: CTCs were isolated from the peripheral blood of two patient cohorts: patients with early-stage breast cancer using immunomagnetic enrichment\/FACS methodology; and patients with late-stage breast cancer using the ANGLE Parsortix microfluidics system. Gene expression profiling using RNA-seq was performed on CTCs and matched primary tumors (PTs) in the early-stage cohort, and on CTCs and matched metastases (METs) for the late-stage cohorts. A quantile normalization approach was used to allow comparison across cohorts and MeCo scores were computed for all samples. The Wilcoxon matched-pairs signed rank test was performed for the comparison of MeCo scores from matching samples within each cohort; the Mann\u2013Whitney unpaired test was used to compare MeCo scores of CTCs across cohorts. Results: In 12 pairs of patients with early-stage breast cancer, MeCo scores in CTCs were significantly higher than in their matched PTs (p = 0.026). Additionally, in 26 pairs of metastatic patient CTCs and METs, MeCo scores were significantly higher in METs compared to matched CTCs (p = 0.0004). MeCo scores of CTCs were similar between patients with early- and late-stage breast cancers, despite differing CTC isolation strategies (epitope-dependent and microfluidics size gradient). Notably, 98% of the genes in the MeCo score were present across evaluable CTC, MET, and PT samples. Conclusions: Our results show that the MeCo score is higher in CTCs than in PTs, and higher in METs compared to CTCs, in early- and late-stage breast cancer, respectively (i.e., PT &lt; CTC &lt; MET). Therefore, the MeCo score is progressively higher throughout the metastatic cascade in breast cancer. These findings demonstrate that mechanical conditioning from primary tumors is retained during metastatic progression, after mechanical induction by ECM stiffness is lost, as cancer cells disseminate through systemic circulation. Additionally, these findings support that cancer cells with higher MeCo scores are more competent with\u2014and potentially selected for\u2014metastatic progression. Importantly, these findings provide a novel feature of CTCs, mechanical conditioning (MeCo), which is associated with higher capacity for metastasis. Furthermore, since the CTC MeCo score is elevated even in early-stage breast cancer, it could provide, in addition to CTC enumeration, a potential prognostic indicator to improve metastatic risk assessment in early disease.<\/jats:p>","DOI":"10.3390\/cancers17101632","type":"journal-article","created":{"date-parts":[[2025,5,12]],"date-time":"2025-05-12T04:21:23Z","timestamp":1747023683000},"page":"1632","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Mechanical Conditioning (MeCo) Score Progressively Increases Through the Metastatic Cascade in Breast Cancer via Circulating Tumor Cells"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8103-4701","authenticated-orcid":false,"given":"Ghassan","family":"Mouneimne","sequence":"first","affiliation":[{"name":"University of Arizona Cancer Center, University of Arizona, Tucson, AZ 85724, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1701-8771","authenticated-orcid":false,"given":"Casey","family":"Connors","sequence":"additional","affiliation":[{"name":"Division of Breast Surgery, Cleveland Clinic, Lerner College of Medicine, 9500 Euclid Ave., Cleveland, OH 44195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam","family":"Watson","sequence":"additional","affiliation":[{"name":"MeCo Diagnostics, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam","family":"Grant","sequence":"additional","affiliation":[{"name":"MeCo Diagnostics, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Campo","sequence":"additional","affiliation":[{"name":"Norris Comprehensive Cancer Center, University of Southern California, 1441 Eastlake Ave., Los Angeles, CA 90033, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0092-1323","authenticated-orcid":false,"given":"Alexander","family":"Ring","sequence":"additional","affiliation":[{"name":"Norris Comprehensive Cancer Center, University of Southern California, 1441 Eastlake Ave., Los Angeles, CA 90033, USA"},{"name":"Department of Medical Oncology and Hematology, University Hospital Z\u00fcrich, R\u00e4mistrasse 100, 8091 Zurich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8780-2951","authenticated-orcid":false,"given":"Pushpinder","family":"Kaur","sequence":"additional","affiliation":[{"name":"Norris Comprehensive Cancer Center, University of Southern California, 1441 Eastlake Ave., Los Angeles, CA 90033, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7534-6124","authenticated-orcid":false,"given":"Julie E.","family":"Lang","sequence":"additional","affiliation":[{"name":"Division of Breast Surgery, Cleveland Clinic, Lerner College of Medicine, 9500 Euclid Ave., Cleveland, OH 44195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1038\/s41568-023-00656-5","article-title":"Linking cell mechanical memory and cancer metastasis","volume":"24","author":"Cambria","year":"2024","journal-title":"Nat. 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