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Liquid biopsy of circulating tumor cells ( CTC ) and circulating tumor  DNA  (ct DNA ) is gaining attention as a method for real‐time monitoring in cancer patients. Conventional methods based upon epithelial cell adhesion molecule (Ep CAM ) expression have a risk of missing the most aggressive  CTC  subpopulations due to epithelial‐mesenchymal transition and may, thus, underestimate the total number of actual  CTC  present in the bloodstream. Techniques utilizing a label‐free inertial microfluidics approach ( LFIMA ) enable efficient capture of  CTC  without the need for Ep CAM  expression. In this study, we optimized a method for analyzing genetic alterations using next‐generation sequencing ( NGS ) of extracted ct DNA  and  CTC  enriched using an  LFIMA  as a first‐phase examination of 30 patients with head and neck cancer, esophageal cancer, gastric cancer and colorectal cancer ( CRC ). Seven patients with advanced  CRC  were enrolled in the second‐phase examination to monitor the emergence of alterations occurring during treatment with epidermal growth factor receptor ( EGFR )‐specific antibodies. Using  LFIMA , we effectively captured  CTC  (median number of  CTC , 14.5 cells/mL) from several types of cancer and detected missense mutations via  NGS  of  CTC  and ct DNA . We also detected time‐dependent genetic alterations that appeared during anti– EGFR  therapy in  CTC  and ct DNA  from  CRC  patients. The results of  NGS  analyses indicated that alterations in the genomic profile revealed by the liquid biopsy could be expanded by using a combination of assays with  CTC  and ct DNA . The study was registered with the University Hospital Medical Information Network Clinical Trials Registry ( ID :  UMIN 000014095).

Monitoring of cancer patients via next‐generation sequencing of patient‐derived circulating tumor cells and tumor DNA