Related Experiment Video
Updated: Jul 19, 2025
Clinical Microfluidic Chip Platform for the Isolation of Versatile Circulating Tumor Cells
Published on: October 13, 2023
Functional Analysis of Viable Circulating Tumor Cells from Triple-Negative Breast Cancer Patients Using TetherChip
Vasileios Vardas1, Julia A Ju2, Athina Christopoulou3
1Laboratory of Biochemistry/Metastatic Signaling, Section of Genetics, Cell Biology and Development, Department of Biology, University of Patras, GR-26504 Patras, Greece.
Abstract:
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 α-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.
Insights
This study isolated viable circulating tumor cells (CTCs) from triple-negative breast cancer (TNBC) patients. A novel method using TetherChip technology effectively evaluated drug responses, showing vinorelbine induces apoptosis and reduces metastasis markers in CTCs.
Area of Science:
- Oncology
- Cell Biology
- Translational Medicine
Background:
- Metastasis is the primary cause of breast cancer mortality, with triple-negative breast cancer (TNBC) being particularly aggressive.
- Microtentacles (McTNs) are crucial for metastasis, and TNBC lacks effective targeted therapies.
- Isolating and functionally analyzing viable circulating tumor cells (CTCs) is essential for personalized treatment strategies.
Purpose of the Study:
- To isolate viable CTCs from TNBC patients for functional drug analysis.
- To evaluate the efficacy of vinorelbine in inducing apoptosis and reducing metastasis-associated markers in CTCs.
- To establish a patient-individualized method for targeting metastatic dissemination.
Main Methods:
- CTCs were isolated from 20 TNBC patients using TetherChip technology and cultured for 5 days.
- Biomarker expression (Detyrosinated α-tubulin, PD-L1) was assessed via immunofluorescence and VyCap analysis.
- Vinorelbine-induced apoptosis was quantified by M30-positive cell detection.
Main Results:
- TetherChips significantly increased CTC yield compared to cytospins (p = 0.006), enabling drug evaluation.
- One-hour vinorelbine treatment induced significant apoptosis in live CTCs (p = 0.010).
- Vinorelbine significantly reduced Detyrosinated α-tubulin and PD-L1 expression (p < 0.001) and disrupted McTNs.
Conclusions:
- TetherChip technology provides a viable protocol for functional CTC analysis and drug efficacy assessment.
- This approach offers critical insights for developing individualized therapies against metastatic breast cancer.
- Targeting metastatic dissemination at a patient-specific level is achievable with this methodology.
