Succinct workflows for circulating tumor cells after enrichment: From systematic counting to mutational profiling

Area of Science:

• Biotechnology
• Oncology
• Molecular Biology

Background:

• Circulating tumor cells (CTCs) are crucial biomarkers for cancer detection and monitoring.
• Current workflows for CTC analysis face challenges in adaptability and integration for genetic studies.

Purpose of the Study:

• To establish a highly adaptable workflow for CTC enumeration and genetic discovery downstream of microfluidic enrichment.
• To bridge the gap between microfluidic enrichment and subsequent CTC analyses for broader clinical and academic applications.

Main Methods:

• Developed a CK/EPCAM-combined immunostaining strategy and an automated analytical pipeline for CTC enumeration.
• Integrated CTC enumeration with next-generation sequencing for genetic analysis using a novel amplicon library.
• Conducted a pilot study involving 56 cancer patients and 21 healthy individuals using a high-throughput microfluidic chip system.

Main Results:

• The workflow demonstrated 80.4% sensitivity and 85.7% specificity for CTC detection (AUC = 0.87).
• CTCs (CK/EpCAM+CD45-) were identified as significant and independent biomarkers for cancer patients (p < 0.01).
• Serial CTC monitoring showed reduced numbers post-treatment, indicating clinical utility; somatic mutations in TP53 and ESR1 were identified in CTCs.

Conclusions:

• The developed workflow provides a systematic and adaptable approach for CTC analysis.
• This integrated workflow facilitates broader applications of CTCs in clinical settings and academic research.
• The study highlights the potential of CTCs for biomarker discovery and pharmacodynamic studies.