부산대학교 도서관

ABSTRACT: Several hundred clinical trials currently explore the role of circulating tumor cell (CTC) analysis for therapy decisions, but assays are lacking for comprehensive molecular characterization of CTCs with diagnostic precision. We therefore combined a workflow for enrichment and isolation of pure CTCs with a non-random whole genome amplification method for single cells and applied it to 510 single CTCs and 189 leukocytes of 66 CTC-positive breast cancer patients. We defined a genome integrity index (GII) to identify single cells suited for molecular characterization by different molecular assays, such as diagnostic profiling of point mutations, gene amplifications and whole genomes of single cells. The reliability of > 90% for successful molecular analysis of high-quality clinical samples selected by the GII enabled assessing the molecular heterogeneity of single CTCs of metastatic breast cancer patients. We readily identified genomic disparity of potentially high relevance between primary tumors and CTCs. Microheterogeneity analysis among individual CTCs uncovered pre-existing cells resistant to ERBB2-targeted therapies suggesting ongoing microevolution at late-stage disease whose exploration may provide essential information for personalized treatment decisions and shed light into mechanisms of acquired drug resistance. SYNOPSIS: (Figure is included in full-text article.)A novel workflow enabling detection, isolation and characterization of single circulating tumors cells (CTCs) from blood suggests that CTCs may harbor genetic alterations undetectable in the primary tumor and associated with therapy resistance. : A novel workflow enabling detection, isolation and characterization of single circulating tumors cells (CTCs) from blood suggests that CTCs may harbor genetic alterations undetectable in the primary tumor and associated with therapy resistance.(Figure is included in full-text article.)