title: Mutational studies on single circulating tumor cells isolated from the blood of inflammatory breast cancer patients.
abstract: OBJECTIVE
  The molecular characterization of circulating tumor cells (CTCs) is critical to identify the key drivers of cancer metastasis and devising therapeutic approaches, particularly for inflammatory breast cancer (IBC) which is usually diagnosed at advance stages and progresses rapidly.
  
  
  METHODS
  Genomic alterations in tumor tissue samples were studied using Foundation One™. Single CTCs were isolated using CellSearch followed by single-cell isolation by DEPArray™. Samples with 20 or more CTCs were chosen to isolate single CTCs using the DEPArray™.
  
  
  RESULTS
  Genomic alterations were studied in primary tumor or metastatic sites from 32 IBC patients. Genes with high-frequency mutations were as follows: TP53 (69%), RB1 (16%), PIK3CA (13%), and also ErbB2 (3%). At least once during treatment, CTCs were detected in 26 patients with metastatic IBC, in two patients with locally advanced IBC, and four patients had no detectable CTCs. Per 7.5 mL of blood, fifteen patients (47%) had ≥20 CTCs and six of them were chosen at random to isolate single CTCs. These cells were tested for the presence of TP53, RB1, PIK3CA, and/or ErbB2 mutations previously found in matching tissue biopsies. The isolated CTCs showed the same mutations as primary or metastatic tumor samples. Intra-patient CTC heterogeneity was found by the presence of different CTC subclones, with some CTCs harboring different combinations of mutated and wild-type genes.
  
  
  CONCLUSIONS
  Our results indicate that CTCs could represent a non-invasive source of cancer cells from which to determine genetic markers as the disease progresses and identify potential therapeutic targets in IBC patients.
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Bingham
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA. Sandra.Fernandez@fccc.edu.|Protocol Support Laboratory, Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA, 19111, USA. Sandra.Fernandez@fccc.edu.
authors.forename: Sandra V
authors.initials: SV
authors.lastname: Fernandez
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Fittipaldi
authors.affiliation: Cynvenio Biosystems, Westlake Village, CA, USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Dempsey
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.
authors.forename: Karen J
authors.initials: KJ
authors.lastname: Ruth
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.|Robert H Lurie Comprehensive Cancer Center of Northwestern University, Chicago, IL, USA.
authors.forename: Massimo
authors.initials: M
authors.lastname: Cristofanilli
authors.affiliation: Fox Chase Cancer Center, Philadelphia, PA, 19111, USA. R.Alpaugh@fccc.edu.|Protocol Support Laboratory, Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA, 19111, USA. R.Alpaugh@fccc.edu.
authors.forename: R
authors.initials: R
authors.lastname: Katherine Alpaugh
mesh_terms: D000328:Adult; D000368:Aged; D001483:Base Sequence; D058534:Class I Phosphatidylinositol 3-Kinases / Q000235:genetics; D004252:DNA Mutational Analysis; D005260:Female; D056726:Genetic Association Studies; D006801:Humans; D058922:Inflammatory Breast Neoplasms / Q000097:blood* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D056286:Retinoblastoma Binding Proteins / Q000235:genetics; D017384:Sequence Deletion; D059010:Single-Cell Analysis; D016019:Survival Analysis; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000235:genetics
chemical_list: D058534:Class I Phosphatidylinositol 3-Kinases; D018719:Erb-b2 Receptor Tyrosine Kinases; D056286:Retinoblastoma Binding Proteins; D016159:Tumor Suppressor Protein p53; D044767:Ubiquitin-Protein Ligases; C000606393:RB1 protein, human; C495901:TP53 protein, human; C484760:PIK3CA protein, human; C508053:ERBB2 protein, human
keywords: CTC; IBC; Single-cell analysis; Tumor heterogeneity