Crystal digital droplet PCR for detection and quantification of circulating EGFR sensitizing and resistance mutations in advanced non-small cell lung cancer

PLoS One. 2017 Aug 22;12(8):e0183319. doi: 10.1371/journal.pone.0183319. eCollection 2017.

Abstract

Over the past years, targeted therapies using tyrosine kinase inhibitors (TKI) have led to an increase in progression-free survival and response rate for a subgroup of non-small cell lung cancer (NSCLC) patients harbouring specific gene abnormalities compared with chemotherapy. However long-lasting tumor regression is rarely achieved, due to the development of resistant tumoral subclones, which requires alternative therapeutic approaches. Molecular profile at progressive disease is a challenge for making adaptive treatment decisions. The aim of this study was to monitor EGFR-mutant tumors over time based on the quantity of mutant DNA circulating in plasma (ctDNA), comparing two different methods, Crystal™ Digital™ PCR and Massive Parallel Sequencing (MPS). In plasma circulating cell free DNA (cfDNA) of 61 advanced NSCLC patients we found an overall correlation of 78% between mutated allelic fraction measured by Crystal Digital PCR and MPS. 7 additional samples with sensitizing mutations and 4 additional samples with the resistance mutation were detected with Crystal Digital PCR, but not with MPS. Monitoring levels of both mutation types over time showed a correlation between levels and trends of mutated ctDNA detected and clinical assessment of disease for the 6 patients tested. In conclusion, Crystal Digital PCR exhibited good performance for monitoring mutational status in plasma cfDNA, and also appeared as better suited to the detection of known mutations than MPS in terms of features such as time to results.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Carcinoma, Non-Small-Cell Lung / genetics*
  • DNA / blood
  • Drug Resistance, Neoplasm / genetics*
  • ErbB Receptors / genetics*
  • Female
  • Humans
  • Lung Neoplasms / genetics*
  • Male
  • Middle Aged
  • Mutation*
  • Polymerase Chain Reaction / methods*

Substances

  • DNA
  • ErbB Receptors

Grants and funding

JM, BA and MD are employees of Stilla technologies. MD is additionally a member on the board of Stilla Technologies. JM and MD provided technical support for using the Naica system and BA provided technical support for statistical analysis. Stilla technologies provided support in the form of salaries for following authors only JM, BA and MD, and material support by providing consumables (Sapphire chips) and equipment (Naica system) free of charge to Gustave-Roussy for this study. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.