Investigating the Role of CTCs with Stem/EMT-like Features in Metastatic Breast Cancer Patients Treated with Eribulin Mesylate

SIMPLE SUMMARY: Eribulin mesylate, an anti-mitotic drug used for the treatment of metastatic breast cancer (BC), exhibits significant effects on cancer cell migration, invasion, and metastatic seeding in experimental models. Interestingly, eribulin treatment has been shown to target the cancer stem cell (CSC) subsets in vitro and reverse the epithelial-to-mesenchymal transition (EMT) state of BC cells. In the current study, circulating tumor cells (CTCs) identified in the peripheral blood of patients with metastatic BC were analyzed at different time points during eribulin treatment and on disease progression. The results contribute new data on the mechanisms of resistance to eribulin mesylate and the prognostic relevance of CTC analyses for eribulin-treated metastatic BC. ABSTRACT: We herein aimed to assess the effect of eribulin mesylate on the cancer stem cell (CSC)/EMT-like phenotype of CTCs, and to investigate the prognostic role of CTC detection and monitoring for eribulin-treated BC patients. Peripheral blood was obtained at baseline (n = 42 patients) and 8 days after treatment initiation (C1D8: n = 22), and on disease progression (PD: n = 26). PBMCs cytospins were immunofluorescently stained for Cytokeratins/ALDH1/TWIST1/DAPI and analyzed via Ariol microscopy. CTCs were detected in 33.3%, 27.3%, and 23.1% of patients at baseline, C1D8, and PD, respectively. Accordingly, partial-EMT+ CTCs represented 61.3%, 0%, and 37.5% of total CTCs, whereas the CSC-like phenotype was consistently expressed by 87.5%, 75%, and 91.7% of CTCs at the respective time points. Interestingly, the CSC+/partial-EMT+ subset prevailed at baseline, but it was eradicated on C1D8 and resurged again during PD. CTC detection at baseline was associated with reduced PFS (p = 0.007) and OS (p = 0.005), and was an independent risk factor for death (HR: 3.779, p = 0.001; multivariate analysis). The CSC+/partial-EMT+ CTCs emerged as the only subset with adverse prognostic significance, while CTC monitoring during eribulin therapy improved the prediction of disease progression. These results indicate that resistant CTC subsets persevere eribulin treatment and highlight the prognostic implications of CTC analyses for eribulin-treated BC patients.