Isolation and Sequencing of Single Circulating Tumor Cells

Area of Science:

• Oncology
• Genomics
• Biotechnology

Background:

• Single-cell sequencing (SCS) is crucial for understanding cancer heterogeneity, therapy resistance, and metastasis.
• Circulating tumor cells (CTCs) present unique challenges due to their rarity and heterogeneity.
• Analyzing CTCs at the single-cell level is vital for personalized cancer treatment.

Purpose of the Study:

• To develop and present a comprehensive workflow for enriching, detecting, and isolating single non-small-cell lung cancer (NSCLC) CTCs.
• To characterize tumor cell subpopulations resistant to targeted therapy using single-cell analysis.
• To enable robust molecular profiling of rare CTCs for improved cancer diagnostics and therapeutics.

Main Methods:

• Development of an integrated workflow for CTC enrichment, detection, and isolation.
• Application of whole-genome amplification (WGA) for single CTC molecular preparation.
• Utilizing targeted next-generation sequencing (NGS) for somatic variant identification and low-pass whole genome sequencing (LP-WGS) for copy number alteration (CNA) analysis.

Main Results:

• Successful isolation and molecular characterization of individual NSCLC CTCs.
• Identification of specific genetic alterations and copy number variations within CTC subpopulations.
• Insights into the molecular mechanisms underlying targeted therapy resistance in NSCLC.

Conclusions:

• The presented workflow enables detailed molecular analysis of rare CTCs, advancing cancer research.
• This methodology facilitates the characterization of therapy-resistant tumor cell subpopulations.
• The findings contribute to a better understanding of NSCLC heterogeneity and resistance, paving the way for improved targeted therapies.