Functional Analysis of Viable Circulating Tumor Cells from Triple-Negative Breast Cancer Patients Using TetherChip Technology

Metastasis, rather than the growth of the primary tumor, accounts for approximately 90% of breast cancer patient deaths. Microtentacles (McTNs) formation represents an important mechanism of metastasis. Triple-negative breast cancer (TNBC) is the most aggressive subtype with limited targeted therapies. The present study aimed to isolate viable circulating tumor cells (CTCs) and functionally analyze them in response to drug treatment. CTCs from 20 TNBC patients were isolated and maintained in culture for 5 days. Biomarker expression was identified by immunofluorescence staining and VyCap analysis. Vinorelbine-induced apoptosis was evaluated based on the detection of M30-positive cells. Our findings revealed that the CTC absolute number significantly increased using TetherChips analysis compared to the number of CTCs in patients' cytospins (p = 0.006) providing enough tumor cells for drug evaluation. Vinorelbine treatment (1 h) on live CTCs led to a significant induction of apoptosis (p = 0.010). It also caused a significant reduction in Detyrosinated & alpha;-tubulin (GLU), programmed death ligand (PD-L1)-expressing CTCs (p < 0.001), and disruption of McTNs. In conclusion, this pilot study offers a useful protocol using TetherChip technology for functional analysis and evaluation of drug efficacy in live CTCs, providing important information for targeting metastatic dissemination at a patient-individualized level.

Automated summary

Metastasis is the main cause of breast cancer patient deaths, and microtentacles formation plays a crucial role in this process. This study aimed to analyze the function of circulating tumor cells (CTCs) in response to drug treatment in triple-negative breast cancer (TNBC) patients. CTCs were isolated from 20 TNBC patients and cultured, and their biomarker expression and drug response were evaluated. The findings showed that TetherChips analysis increased the number of CTCs, allowing for drug evaluation. Vinorelbine treatment induced apoptosis and reduced specific markers in CTCs. This pilot study provides valuable insights for individualized targeting of metastasis.