Single-cell transcriptomic profiling of platelet-adherent circulating tumor cells using a microcavity-gel...

Area of Science:

• Oncology
• Molecular Biology
• Genomics

Background:

• Pancreatic cancer is highly lethal with poor survival rates due to metastasis and drug resistance.
• Circulating tumor cells (CTCs) are crucial for understanding pancreatic cancer metastasis and treatment challenges.

Purpose of the Study:

• To develop and validate a novel single-cell transcriptome analysis platform for pancreatic cancer CTCs.
• To investigate the biological characteristics and potential therapeutic vulnerabilities of pancreatic CTCs.

Main Methods:

• Utilized a microcavity array (MCA) and gel-based cell manipulation (GCM) for marker-independent CTC recovery.
• Performed high-quality single-cell RNA sequencing on pancreatic cancer cell lines and patient-derived CTCs.
• Conducted spiking experiments to assess platform specificity and minimize blood cell contamination.

Main Results:

• The MCA-GCM platform enabled high-quality single-cell RNA sequencing while preserving intrinsic molecular features.
• Patient-derived CTCs showed significant epithelial-mesenchymal transition (EMT)-like features.
• CTCs highly expressed platelet-related genes (e.g., PF4, PPBP), suggesting platelet-driven EMT, and were arrested in the G1 cell cycle phase.

Conclusions:

• The microcavity-gel manipulation platform is effective for robust single-cell transcriptomic profiling of CTCs.
• Pancreatic CTCs exhibit EMT and platelet interactions, indicating potential mechanisms for metastasis.
• G1 cell cycle arrest in CTCs suggests a survival strategy under therapeutic pressure.