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Int. J. Mol. Sci. 2016, 17(10), 1665; doi:10.3390/ijms17101665

Detection and Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer

1
Department of Medicine-Hematology and Oncology, Robert H Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA
2
ScreenCell SA, Sarcelles 95200, France
3
Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA
4
Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA
5
Department of Molecular Medicine, University of Rome “Sapienza”, Rome 00185, Italy
*
Authors to whom correspondence should be addressed.
Academic Editor: Dario Marchetti
Received: 5 August 2016 / Revised: 22 September 2016 / Accepted: 23 September 2016 / Published: 30 September 2016
(This article belongs to the Special Issue Circulating Tumor Cells)
View Full-Text   |   Download PDF [2582 KB, uploaded 30 September 2016]   |  

Abstract

The availability of blood-based diagnostic testing using a non-invasive technique holds promise for real-time monitoring of disease progression and treatment selection. Circulating tumor cells (CTCs) have been used as a prognostic biomarker for the metastatic breast cancer (MBC). The molecular characterization of CTCs is fundamental to the phenotypic identification of malignant cells and description of the relevant genetic alterations that may change according to disease progression and therapy resistance. However, the molecular characterization of CTCs remains a challenge because of the rarity and heterogeneity of CTCs and technological difficulties in the enrichment, isolation and molecular characterization of CTCs. In this pilot study, we evaluated circulating tumor associated cells in one blood draw by size exclusion technology and cytological analysis. Among 30 prospectively enrolled MBC patients, CTCs, circulating tumor cell clusters (CTC clusters), CTCs of epithelial–mesenchymal transition (EMT) and cancer associated macrophage-like cells (CAMLs) were detected and analyzed. For molecular characterization of CTCs, size-exclusion method for CTC enrichment was tested in combination with DEPArray™ technology, which allows the recovery of single CTCs or pools of CTCs as a pure CTC sample for mutation analysis. Genomic mutations of TP53 and ESR1 were analyzed by targeted sequencing on isolated 7 CTCs from a patient with MBC. The results of genomic analysis showed heterozygous TP53 R248W mutation from one single CTC and pools of three CTCs, and homozygous TP53 R248W mutation from one single CTC and pools of two CTCs. Wild-type ESR1 was detected in the same isolated CTCs. The results of this study reveal that size-exclusion method can be used to enrich and identify circulating tumor associated cells, and enriched CTCs were characterized for genetic alterations in MBC patients, respectively. View Full-Text
Keywords: metastatic breast cancer (MBC); circulating tumor associated cells; circulating tumor cells (CTCs); circulating tumor cell clusters (CTC clusters); epithelial–mesenchymal transition (EMT); cancer associated macrophage-like cells (CAMLs); size-exclusion technology metastatic breast cancer (MBC); circulating tumor associated cells; circulating tumor cells (CTCs); circulating tumor cell clusters (CTC clusters); epithelial–mesenchymal transition (EMT); cancer associated macrophage-like cells (CAMLs); size-exclusion technology
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MDPI and ACS Style

Mu, Z.; Benali-Furet, N.; Uzan, G.; Znaty, A.; Ye, Z.; Paolillo, C.; Wang, C.; Austin, L.; Rossi, G.; Fortina, P.; Yang, H.; Cristofanilli, M. Detection and Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer. Int. J. Mol. Sci. 2016, 17, 1665.

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