Molecular profiling of single circulating tumor cells with diagnostic intention

Bernhard Polzer, Gianni Medoro, Sophie Pasch, Francesca Fontana, Laura Zorzino, Aurelia Pestka, Ulrich Andergassen, Franziska Meier-Stiegen, Zbigniew T. Czyz, Barbara Alberter, Steffi Treitschke, Thomas Schamberger, Maximilian Sergio, Giulia Bregola, Anna Doffini, Stefano Gianni, Alex Calanca, Giulio Signorini, Chiara Bolognesi, Arndt HartmannPeter A. Fasching, Maria T. Sandri, Brigitte Rack, Tanja Fehm, Giuseppe Giorgini, Nicolò Manaresi, Christoph A. Klein

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)1371-1386
Number of pages16
JournalEMBO Molecular Medicine
Issue number11
Publication statusPublished - Nov 1 2014


  • Breast cancer
  • Circulating tumor cells
  • Metastasis
  • Single cell analysis

ASJC Scopus subject areas

  • Molecular Medicine


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