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Detection of Circulating Tumor Cells Using the Attune NxT
Mandy Gruijs1,2,3, Carolien Zeelen1,2,3, Tessa Hellingman2,4
1Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology and Immunology, De Boelelaan 1117, Amsterdam 1081 HV, The Netherlands.
International Journal of Molecular Sciences
|January 8, 2023
Summary
A new flow cytometry method using Attune NxT for detecting circulating tumor cells (CTCs) showed promise but requires morphological confirmation. While easier to use, it may not surpass CellSearch for low-abundance CTC detection in cancer patients.
Area of Science:
- Oncology
- Biotechnology
- Flow Cytometry
Background:
- Circulating tumor cells (CTCs) are biomarkers for solid malignancies, correlating with poor patient survival.
- Current CTC detection methods, like FDA-approved CellSearch, have limitations in sensitivity for certain cancers and early disease stages.
- Existing methods are primarily limited to metastatic colorectal, breast, and prostate cancers.
Purpose of the Study:
- To develop and evaluate a novel flow cytometry-based method, Attune NxT, for detecting rare circulating tumor cells (CTCs) without prior enrichment.
- To assess the efficacy of the Attune NxT system in detecting EpCAM+EGFR+ tumor cells in spiked samples and patient blood.
- To compare the performance of the Attune NxT system with the established CellSearch method for CTC detection.
Main Methods:
- Utilized the Attune NxT flow cytometer, a system designed for rare event detection, for CTC analysis.
- Tested the system's recovery yield and detection range using healthy donor blood spiked with known tumor cell lines (EpCAM+EGFR+).
- Performed morphological confirmation of detected events using the Attune CytPix imaging system.
- Analyzed blood samples from patients with metastatic colorectal and lung cancer.
Main Results:
- The Attune NxT system demonstrated an average recovery yield of 75% for spiked tumor cells, with a detection range down to 10 cells per sample.
- Increased EpCAM+EGFR+ events were detected in over half of the analyzed patient samples (colorectal and lung cancer).
- However, most detected events lacked tumor cell-like morphology, and the Attune NxT did not prove superior to CellSearch in detecting low CTC numbers, highlighting the need for morphological validation.
Conclusions:
- The Attune NxT system offers a user-friendly approach for rare event detection in liquid biopsies.
- Morphological confirmation is critical to differentiate true CTCs from false positives, especially when low cell counts are involved.
- The Attune NxT system may not offer a significant advantage over CellSearch for detecting low-abundance CTCs, despite its easier sample handling and analysis.
