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Detection of Targetable Alterations in Non-small Cell Lung Cancer using Next-generation Sequencing
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Detection of Targetable Alterations in Non-small Cell Lung Cancer using Next-generation Sequencing

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Development, validation, and comparison of gene analysis methods for detecting EGFR mutation from non-small cell lung

Masaki Hanibuchi1,2, Akira Kanoh3, Takuya Kuramoto3

  • 1Department of Respiratory Medicine and Rheumatology, Graduate School of Biomedical Sciences, Tokushima University, Tokushima, 770-8503, Japan.

Oncotarget
|June 21, 2019
PubMed

Insights

Circulating free DNA (cfDNA) assays show clinical relevance for detecting acquired epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. These highly sensitive cfDNA tests can identify T790M mutations after EGFR-tyrosine kinase inhibitor (TKI) treatment failure.

Area of Science:

  • Molecular Oncology
  • Genetics
  • Cancer Research

Background:

  • Second-line epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) treatment efficacy in non-small cell lung cancer (NSCLC) is often limited by acquired resistance mutations, such as T790M.
  • Detecting these resistance mutations is crucial for guiding subsequent treatment strategies, but tissue biopsies can be invasive and may not always be feasible.
  • Circulating free DNA (cfDNA) analysis offers a less invasive alternative for molecular profiling, yet its sensitivity and clinical utility in this context require further elucidation.

Purpose of the Study:

  • To evaluate the feasibility and sensitivity of novel circulating free DNA (cfDNA)-based detection methods for epidermal growth factor receptor (EGFR) mutations, including T790M, in patients with non-small cell lung cancer (NSCLC) undergoing second-line EGFR-TKI treatment.
  • To assess the clinical usability and concordance of cfDNA-based assays compared to traditional tissue biopsy diagnoses for identifying acquired EGFR mutations.

Main Methods:

  • Developed and employed three highly sensitive, novel assays for detecting EGFR mutations, including T790M, in cfDNA extracted from plasma samples of 45 NSCLC patients with known activating EGFR mutations.
  • Compared the results of cfDNA-based mutation detection with definitive tissue-based diagnoses, including re-biopsy data where available.
  • Calculated specificity, sensitivity, and concordance rates for the cfDNA assays against tissue-based results.

Main Results:

  • The cfDNA-based methods demonstrated high specificity (96-100%) with a low pseudo-positive rate, while sensitivity ranged from 56-67%.
  • In patients who failed prior EGFR-TKI treatment, T790M mutations were detected in 41-46% of cfDNA samples, comparable to literature values for tissue-based re-biopsies.
  • Concordance rates for specific EGFR mutations between cfDNA and tissue ranged from 83-95%, with overall agreement in T790M detection between 50-63% and activating mutations between 63-100% in a subset of eight patients.

Conclusions:

  • Newly developed cfDNA assays possess sufficient sensitivity to detect circulating tumor DNA, demonstrating clinical relevance for identifying acquired EGFR mutations, particularly T790M, in NSCLC patients post-EGFR-TKI failure.
  • cfDNA-based mutation detection presents a viable, less invasive approach for monitoring treatment response and guiding therapeutic decisions in NSCLC.
  • The high specificity and comparable detection rates for T790M suggest that cfDNA analysis can complement or potentially replace re-biopsies in certain clinical scenarios.
Keywords:
circulating free DNAepidermal growth factor receptor mutationepidermal growth factor receptor-tyrosine kinase inhibitornon-small cell lung cancer

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