title: Detection and Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer.
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 ofTP53andESR1were analyzed by targeted sequencing on isolated 7 CTCs from a patient with MBC. The results of genomic analysis showed heterozygousTP53R248W mutation from one single CTC and pools of three CTCs, and homozygousTP53R248W mutation from one single CTC and pools of two CTCs. Wild-typeESR1was 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.
authors.affiliation: Department of Medicine-Hematology and Oncology, Robert H Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA. zhaomei.mu@northwestern.edu.
authors.forename: Zhaomei
authors.initials: Z
authors.lastname: Mu
authors.affiliation: ScreenCell SA, Sarcelles 95200, France. benali@screencell.com.
authors.forename: Naoual
authors.initials: N
authors.lastname: Benali-Furet
authors.affiliation: ScreenCell SA, Sarcelles 95200, France. guzan@screencell.com.
authors.forename: Georges
authors.initials: G
authors.lastname: Uzan
authors.affiliation: ScreenCell SA, Sarcelles 95200, France. aznaty@screencell.com.
authors.forename: Anaëlle
authors.initials: A
authors.lastname: Znaty
authors.affiliation: Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. Zhong.Ye@jefferson.edu.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Ye
authors.affiliation: Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. carmela.Paolillo@jefferson.edu.
authors.forename: Carmela
authors.initials: C
authors.lastname: Paolillo
authors.affiliation: Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. Chun.Wang@jefferson.edu.
authors.forename: Chun
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. laustin@gmail.com.
authors.forename: Laura
authors.initials: L
authors.lastname: Austin
authors.affiliation: Department of Medicine-Hematology and Oncology, Robert H Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA. giovirossi85@yahoo.it.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Rossi
authors.affiliation: Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. paolo.Fortina@jefferson.edu.|Department of Molecular Medicine, University of Rome "Sapienza", Rome 00185, Italy. paolo.Fortina@jefferson.edu.
authors.forename: Paolo
authors.initials: P
authors.lastname: Fortina
authors.affiliation: Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. hushan.yang@jefferson.edu.
authors.forename: Hushan
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Medicine-Hematology and Oncology, Robert H Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA. massimo.cristofanilli@nm.org.
authors.forename: Massimo
authors.initials: M
authors.lastname: Cristofanilli
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000378:metabolism / Q000473:pathology*; D004252:DNA Mutational Analysis; D058750:Epithelial-Mesenchymal Transition; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D008856:Microscopy, Fluorescence; D009154:Mutation; D009362:Neoplasm Metastasis; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D016159:Tumor Suppressor Protein p53
keywords: cancer associated macrophage-like cells (CAMLs); circulating tumor associated cells; circulating tumor cell clusters (CTC clusters); circulating tumor cells (CTCs); epithelial–mesenchymal transition (EMT); metastatic breast cancer (MBC); size-exclusion technology