Evaluating the Genomic Yield of a Single Endobronchial Ultrasound-guided Transbronchial Needle Aspiration in Lung Cancer: Meeting the Challenge of Doing More With Less

Clin Lung Cancer. 2017 Nov;18(6):e467-e472. doi: 10.1016/j.cllc.2017.05.006. Epub 2017 May 10.

Abstract

Background: Minimally invasive techniques, including endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA), yield small specimens that are adequate for cytologic diagnosis of lung cancer, but also need to provide material for molecular analysis to guide treatment. The number of EBUS-TBNA passes needed for mutation testing remains unclear. We sought to assess the adequacy of a single pass for genomic profiling of actionable mutations.

Methods: In a prospective observational study, paired samples from the same lesion were obtained from patients undergoing EBUS-TBNA for lung cancer diagnosis/staging. Following tumor cell confirmation by rapid on-site evaluation, a "reference" sample comprising ≥ 3 passes was obtained and formalin-fixed paraffin-embedded. A "study" sample comprising a single pass was taken and snap-frozen. The primary outcome was DNA yield and quality from a single pass. The secondary outcome was diagnostic accuracy of a single pass for detecting actionable mutations.

Results: In 40 patients, single-pass specimens yielded a mean 3.98 μg of highly intact DNA, well above the minimum threshold for targeted sequencing, which was performed in adenocarcinoma cases (n = 24). In 23 cases, there was 100% agreement in mutation status between reference and study samples. In 1 case, the reference sample failed to generate a molecular diagnosis owing to insufficient tumor cells; however, the study specimen identified a KRAS mutation. Tumor cell percentage in mutation-positive specimens was 1% to 70%, suggesting that single-pass samples detect mutations even when tumor cell content is low.

Conclusion: Single EBUS-TBNA passes yield DNA of high quantity and quality with high accuracy for molecular profiling, irrespective of tumor cell content.

Keywords: EBUS-TBNA; Genetic analysis; Molecular analysis; Non–small-cell lung cancer; Single pass.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / diagnosis*
  • Adenocarcinoma / genetics
  • Adenocarcinoma / pathology
  • Aged
  • Aged, 80 and over
  • DNA, Neoplasm
  • Endoscopic Ultrasound-Guided Fine Needle Aspiration / methods*
  • Female
  • Genomics / methods*
  • Humans
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques
  • Mutation
  • Neoplasm Staging
  • Prospective Studies
  • Proto-Oncogene Proteins p21(ras) / genetics

Substances

  • DNA, Neoplasm
  • KRAS protein, human
  • Proto-Oncogene Proteins p21(ras)