title: TP53 mutations detected in circulating tumor cells present in the blood of metastatic triple negative breast cancer patients.
abstract: BACKGROUND
  Circulating tumor cells (CTCs) are tumor cells shed from either primary tumors or its metastases that circulate in the peripheral blood of patients with metastatic cancers. The molecular characterization of the CTCs is critical to identifying the key drivers of cancer metastasis and devising therapeutic approaches. However, the molecular characterization of CTCs is difficult to achieve because their isolation is a major technological challenge.
  
  
  METHODS
  CTCs from two triple negative breast cancer patients were enriched using CellSearch and single cells selected by DEPArray™. A TP53 R110 fs*13 mutation identified by next generation sequencing in the breast and chest skin biopsies of both patients was studied in single CTCs.
  
  
  RESULTS
  From 6 single CTC isolated from one patient, 1 CTC had TP53 R110 delC, 1 CTC showed the TP53 R110 delG mutation, and the remaining 4 single CTCs showed the wild type p53 sequence; a pool of 14 CTCs isolated from the same patient also showed TP53 R110 delC mutation. In the tumor breast tissue of this patient, only the TP53 R110 delG mutation was detected. In the second patient a TP53 R110 delC mutation was detected in the chest wall skin biopsy; from the peripheral blood of this patient, 5 single CTC and 6 clusters of 2 to 6 CTCs were isolated; 3 of the 5 single CTCs showed the TP53 R110 delC mutation and 2 CTCs showed the wild type TP53 allele; from the clusters, 5 showed the TP53 R110 delC mutation, and 1 cluster the wild type TP53 allele. Single white blood cells isolated as controls from both patients only showed the wild type TP53 allele.
  
  
  CONCLUSIONS
  We are able to isolate uncontaminated CTCs and achieve single cell molecular analysis. Our studies showed the presence of different CTC sub-clones in patients with metastatic breast cancer. Some CTCs had the same TP53 mutation as their matching tumor samples although others showed either a different TP53 mutation or the wild type allele. Our results indicate that CTCs could represent a non-invasive source of cancer cells from which to determine genetic markers of the disease progression and potential therapeutic targets.
authors.forename: Sandra V
authors.initials: SV
authors.lastname: Fernandez
authors.forename: Catherine
authors.initials: C
authors.lastname: Bingham
authors.forename: Patricia
authors.initials: P
authors.lastname: Fittipaldi
authors.forename: Laura
authors.initials: L
authors.lastname: Austin
authors.forename: Juan
authors.initials: J
authors.lastname: Palazzo
authors.forename: Gary
authors.initials: G
authors.lastname: Palmer
authors.forename: Katherine
authors.initials: K
authors.lastname: Alpaugh
authors.forename: Massimo
authors.initials: M
authors.lastname: Cristofanilli
mesh_terms: D001483:Base Sequence; D018270:Carcinoma, Ductal, Breast; D004252:DNA Mutational Analysis; D005260:Female; D006801:Humans; D058922:Inflammatory Breast Neoplasms; D008969:Molecular Sequence Data; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D017384:Sequence Deletion; D064726:Triple Negative Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53