Enumeration and targeted analysis of KRAS, BRAF and PIK3CA mutations in CTCs captured by a label-free platform: Comparison to ctDNA and tissue in metastatic colorectal cancer

Oncotarget. 2016 Dec 20;7(51):85349-85364. doi: 10.18632/oncotarget.13350.

Abstract

Treatment of advanced colorectal cancer (CRC) requires multimodal therapeutic approaches and need for monitoring tumor plasticity. Liquid biopsy biomarkers, including CTCs and ctDNA, hold promise for evaluating treatment response in real-time and guiding therapeutic modifications. From 15 patients with advanced CRC undergoing liver metastasectomy with curative intent, we collected 41 blood samples at different time points before and after surgery for CTC isolation and quantification using label-free Vortex technology. For mutational profiling, KRAS, BRAF, and PIK3CA hotspot mutations were analyzed in CTCs and ctDNA from 23 samples, nine matched liver metastases and three primary tumor samples. Mutational patterns were compared. 80% of patient blood samples were positive for CTCs, using a healthy baseline value as threshold (0.4 CTCs/mL), and 81.4% of captured cells were EpCAM+ CTCs. At least one mutation was detected in 78% of our blood samples. Among 23 matched CTC and ctDNA samples, we found a concordance of 78.2% for KRAS, 73.9% for BRAF and 91.3% for PIK3CA mutations. In several cases, CTCs exhibited a mutation that was not detected in ctDNA, and vice versa. Complementary assessment of both CTCs and ctDNA appears advantageous to assess dynamic tumor profiles.

Keywords: Vortex; circulating tumor DNA; circulating tumor cells; colorectal cancer; liquid biopsy.

Publication types

  • Comparative Study

MeSH terms

  • Biomarkers, Tumor / blood
  • Biomarkers, Tumor / genetics*
  • Circulating Tumor DNA / blood
  • Circulating Tumor DNA / genetics*
  • Class I Phosphatidylinositol 3-Kinases / blood
  • Class I Phosphatidylinositol 3-Kinases / genetics*
  • Colorectal Neoplasms / blood
  • Colorectal Neoplasms / enzymology
  • Colorectal Neoplasms / genetics*
  • Colorectal Neoplasms / pathology
  • DNA Mutational Analysis / instrumentation
  • DNA Mutational Analysis / methods*
  • Genetic Predisposition to Disease
  • HCT116 Cells
  • Humans
  • Lab-On-A-Chip Devices
  • Microfluidic Analytical Techniques
  • Mutation*
  • Neoplastic Cells, Circulating / metabolism*
  • Neoplastic Cells, Circulating / pathology
  • Phenotype
  • Predictive Value of Tests
  • Prognosis
  • Proto-Oncogene Proteins B-raf / blood
  • Proto-Oncogene Proteins B-raf / genetics*
  • Proto-Oncogene Proteins p21(ras) / blood
  • Proto-Oncogene Proteins p21(ras) / genetics*
  • Reproducibility of Results
  • Time Factors

Substances

  • Biomarkers, Tumor
  • Circulating Tumor DNA
  • KRAS protein, human
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • Proto-Oncogene Proteins p21(ras)