Research Papers:

Mutational analysis of single circulating tumor cells by next generation sequencing in metastatic breast cancer

Francesca De Luca, Giada Rotunno, Francesca Salvianti, Francesca Galardi, Marta Pestrin, Stefano Gabellini, Lisa Simi, Irene Mancini, Alessandro Maria Vannucchi, Mario Pazzagli, Angelo Di Leo and Pamela Pinzani _

PDF  |  HTML  |  How to cite

Oncotarget. 2016; 7:26107-26119. https://doi.org/10.18632/oncotarget.8431

Metrics: PDF 4062 views  |   HTML 4789 views  |   ?  


Francesca De Luca1,*, Giada Rotunno2,*, Francesca Salvianti3,*, Francesca Galardi1, Marta Pestrin1, Stefano Gabellini1, Lisa Simi3, Irene Mancini3, Alessandro Maria Vannucchi2, Mario Pazzagli3, Angelo Di Leo1, Pamela Pinzani3

1Sandro Pitigliani Medical Oncology Department, Hospital of Prato, Istituto Toscano Tumori, 59100, Prato, Italy

2Department of Experimental and Clinical Medicine, University of Florence, 50139, Florence, Italy

3Department of Clinical, Experimental and Biomedical Sciences, University of Florence, 50139, Florence, Italy

*These authors contributed equally to this work

Correspondence to:

Pamela Pinzani, email: [email protected]

Keywords: breast cancer, circulating tumor cells, next generation sequencing, single cell sequencing, somatic mutations

Received: December 21, 2015     Accepted: March 07, 2016     Published: March 28, 2016


Circulating Tumor Cells (CTCs) represent a “liquid biopsy” of the tumor potentially allowing real-time monitoring of cancer biology and therapies in individual patients.

The purpose of the study was to explore the applicability of a protocol for the molecular characterization of single CTCs by Next Generation Sequencing (NGS) in order to investigate cell heterogeneity and provide a tool for a personalized medicine approach.

CTCs were enriched and enumerated by CellSearch in blood from four metastatic breast cancer patients and singularly isolated by DEPArray. Upon whole genome amplification 3–5 single CTCs per patient were analyzed by NGS for 50 cancer-related genes.

We found 51 sequence variants in 25 genes. We observed inter- and intra-patient heterogeneity in the mutational status of CTCs.

The highest number of somatic deleterious mutations was found in the gene TP53, whose mutation is associated with adverse prognosis in breast cancer.

The discordance between the mutational status of the primary tumor and CTCs observed in 3 patients suggests that, in advanced stages of cancer, CTC characteristics are more closely linked to the dynamic modifications of the disease status.

In one patient the mutational profiles of CTCs before and during treatment shared only few sequence variants.

This study supports the applicability of a non-invasive approach based on the liquid biopsy in metastatic breast cancer patients which, in perspective, should allow investigating the clonal evolution of the tumor for the development of new therapeutic strategies in precision medicine.

Creative Commons License All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 4.0 License.
PII: 8431