MINDEC-An Enhanced Negative Depletion Strategy for Circulating Tumour Cell Enrichment

Sci Rep. 2016 Jul 19:6:28929. doi: 10.1038/srep28929.

Abstract

Most current methods of circulating tumour cell (CTC) enrichment target the epithelial protein EpCAM, which is commonly expressed in adenocarcinoma cells. However, such methods will not recover the fraction of CTCs that have a non-epithelial phenotype due to epithelial-mesenchymal transition. For phenotype-independent CTC enrichment, we developed a new enhanced negative depletion strategy-termed MINDEC-that is based on multi-marker (CD45, CD16, CD19, CD163, and CD235a/GYPA) depletion of blood cells rather than targeted enrichment of CTCs. Here we validated the performance of MINDEC using epithelial and mesenchymal cancer cell lines, demonstrating a mean recovery of 82 ± 10%, high depletion (437 ± 350 residual white blood cells (WBCs)/mL peripheral blood), linearity between spiked and recovered cells (correlation coefficient: r = 0.995), and a low detection limit (≥1 cell recovered in all four replicates spiked with 3 cells). For clinical validation of this method, we enumerated CTCs in peripheral blood samples from patients with metastatic pancreatic cancer, detecting CTCs in 15 of 21 blood samples (71%) from 9 patients. The promising performance of the MINDEC enrichment strategy in our study encourages validation in larger clinical trials.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / blood
  • Antigens, CD19 / blood
  • Antigens, Differentiation, Myelomonocytic / blood
  • Antigens, Neoplasm / blood*
  • Biomarkers, Tumor / blood
  • Breast Neoplasms / blood
  • Cell Adhesion Molecules / blood*
  • Cell Line, Tumor
  • Cell Separation / methods
  • Cytological Techniques*
  • Epithelial Cell Adhesion Molecule / metabolism*
  • Epithelial-Mesenchymal Transition
  • Female
  • GPI-Linked Proteins / blood
  • Glycophorins / blood
  • Humans
  • Leukocyte Common Antigens / blood
  • Limit of Detection
  • Lung Neoplasms / blood
  • MCF-7 Cells
  • Mesothelioma / blood
  • Neoplastic Cells, Circulating / pathology*
  • Pancreatic Neoplasms / blood
  • Phenotype
  • Receptors, Cell Surface / blood
  • Receptors, IgG / blood
  • Reproducibility of Results

Substances

  • Antigens, CD
  • Antigens, CD19
  • Antigens, Differentiation, Myelomonocytic
  • Antigens, Neoplasm
  • Biomarkers, Tumor
  • CD163 antigen
  • CD19 molecule, human
  • Cell Adhesion Molecules
  • EPCAM protein, human
  • Epithelial Cell Adhesion Molecule
  • FCGR3B protein, human
  • GPI-Linked Proteins
  • GYPA protein, human
  • Glycophorins
  • Receptors, Cell Surface
  • Receptors, IgG
  • Leukocyte Common Antigens
  • PTPRC protein, human