1. Molecular profiling of single circulating tumor cells with diagnostic intention
- Author
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Stefano Gianni, Anna Doffini, Brigitte Rack, Laura Zorzino, Ulrich Andergassen, Giuseppe Giorgini, Giulio Signorini, Franziska Meier-Stiegen, Tanja Fehm, Giulia Bregola, Bernhard Polzer, Aurelia Pestka, Francesca Fontana, Alex Calanca, Gianni Medoro, Sophie Pasch, Steffi Treitschke, Arndt Hartmann, Chiara Bolognesi, Thomas Schamberger, Peter A. Fasching, Maria T. Sandri, M. Sergio, Zbigniew T. Czyż, Nicolò Manaresi, Christoph Klein, Barbara Alberter, and Publica
- Subjects
breast - cancer ,single cell analysis ,Genomics ,Breast Neoplasms ,Computational biology ,Biology ,circulating tumor cells ,Bioinformatics ,Genome ,Metastasis ,Breast cancer ,Circulating tumor cell ,breast cancer ,Single-cell analysis ,medicine ,metastasis ,Humans ,Pathology, Molecular ,Research Articles ,Whole Genome Amplification ,medicine.disease ,Neoplastic Cells, Circulating ,Metastatic breast cancer ,Molecular Medicine ,Female ,Single-Cell Analysis - 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.
- Published
- 2014