Efficient utilization of EBUS-TBNA samples for both diagnosis and molecular analyses

F Oezkan, Am Khan, P Zarogoulidis, W Hohenforst-Schmidt, D Theegarten, K Yasufuku, T Nakajima, L Freitag, K Darwiche, F Oezkan, Am Khan, P Zarogoulidis, W Hohenforst-Schmidt, D Theegarten, K Yasufuku, T Nakajima, L Freitag, K Darwiche

Abstract

Background: The assessment of an increasing number of molecular markers is becoming a standard requirement from endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) specimens. However, it is unclear how many needle passes should be performed and the amount of lung cancer cells that should be sent for molecular analyses. The objective of this study was to determine if it is feasible to divide the material obtained by EBUS-TBNA to allow for molecular analysis without compromising the accuracy of mediastinal staging.

Objective: We aimed to determine if dividing EBUS-TBNA specimens has a negative impact on either histopathological diagnosis or molecular analysis.

Methods: EBUS-TBNA was performed in 249 enlarged lymph nodes. Negative or ambiguous histopathological results were confirmed by surgical means and clinical follow-up over 6 months. The tissue obtained by EBUS-TBNA was placed onto a glass slide and divided for histopathological workup and molecular analysis. The number of passes was recorded. Both the accuracy of the mediastinal lymph node staging and the applicability of the sample division for molecular analysis were assessed.

Results: Each lymph node was punctured an average of 3.18 times and division of the obtained material for diagnosis and molecular analysis was feasible in all cases. The sensitivity and accuracy of the mediastinal lymph node staging were 96.6% and 97.6%, respectively. A cytokeratin (CK)-19-mRNA concentration-based molecular test was feasible in 74.1% of cases.

Conclusion: Dividing EBUS-TBNA samples for both histopathological diagnosis and molecular testing is feasible and does not compromise the accuracy of mediastinal staging. This method may be an alternative to taking additional needle passes for molecular analyses.

Keywords: CK-19-mRNA; lung cancer; lymph node sampling; molecular marker.

Figures

Figure 1
Figure 1
Flowchart of the lymph nodes evaluated in the study (CONSORT). Abbreviations: EBUS-TBNA, endobronchial ultrasound-guided transbronchial needle aspiration; LN, lymph nodes.
Figure 2
Figure 2
EBUS-TBNA material is placed onto a glass slide, mixed, and divided into two equivalent parts and sent for histopathological and molecular analysis, respectively. The glass slide is then referred to fast track cytological evaluation. Abbreviation: EBUS-TBNA, endobronchial ultrasound-guided transbronchial needle aspiration.

References

    1. Varela-Lema L, Fernández-Villar A, Ruano-Ravina A. Effectiveness and safety of endobronchial ultrasound-transbronchial needle aspiration: a systematic review. Eur Respir J. 2009;33(5):1156–1164.
    1. Herth FJ, Eberhardt R, Vilmann P, Krasnik M, Ernst Al. Real-time endobronchial ultrasound guided transbronchial needle aspiration for sampling mediastinal lymph nodes. Thorax. 2006;61(9):795–798.
    1. Yasufuku K, Pierre A, Darling G, et al. A prospective controlled trial of endobronchial ultrasound-guided transbronchial needle aspiration compared with mediastinoscopy for mediastinal lymph node staging of lung cancer. J Thorac Cardiovasc Surg. 2011;142(6):1393–1400. e1.
    1. Nakajima T, Yasufuku K. How I do it – optimal methodology for multidirectional analysis of endobronchial ultrasound-guided transbronchial needle aspiration samples. J Thorac Oncol. 2011;6(1):203–206.
    1. Wallace MB, Block MI, Gillanders W, et al. Accurate molecular detection of non-small cell lung cancer metastases in mediastinal lymph nodes sampled by endoscopic ultrasound-guided needle aspiration. Chest. 2005;127(2):430–437.
    1. Bugalho A, Martins C, Dias SS, et al. Cytokeratin 19, carcinoembryonic antigen, and epithelial cell adhesion molecule detect lung cancer lymph node metastasis in endobronchial ultrasound-guided tranbronchial aspiration samples. Clin Lung Cancer. 2013;14(6):704–712.
    1. Lawson MH, Rassl DM, Cummings NM, et al. Tissue banking of diagnostic lung cancer biopsies for extraction of high quality RNA. J Thorac Oncol. 2010;5(7):956–963.
    1. Nana-Sinkam SP, Powell CA. Molecular biology of lung cancer: Diagnosis and management of lung cancer, 3rd ed: American College of Chest Physicians evidence-based clinical practice guidelines. Chest. 2013;143(Suppl 5):e30S–e39S.
    1. Wiesweg M, Ting S, Reis H, et al. Feasibility of preemptive biomarker profiling for personalised early clinical drug development at a Comprehensive Cancer Center. Eur J Cancer. 2013;49(15):3076–3082.
    1. Darwiche K, Zarogoulidis P, Baehner K, et al. Assessment of SHOX2 methylation in EBUS-TBNA specimen improves accuracy in lung cancer staging. Ann Oncol. 2013;24(11):2866–2870.
    1. Brustugun OT, Khattak AM, Trømborg AK, et al. BRAF-mutations in non-small cell lung cancer. Lung Cancer. 2014;84(1):36–38.
    1. Scarpa A, Sikora K, Fassan M, et al. Molecular typing of lung adenocarcinoma on cytological samples using a multigene next generation sequencing panel. PLoS One. 2013;8(11):e80478.
    1. Herbst RS, Lippman SM. Molecular signatures of lung cancer – toward personalized therapy. N Engl J Med. 2007;356(1):76–78.
    1. Santis G, Angell R, Nickless G, et al. Screening for EGFR and KRAS mutations in endobronchial ultrasound derived transbronchial needle aspirates in non-small cell lung cancer using COLD-PCR. PLoS One. 2011;6(9):e25191.
    1. Nakajima T, Kimura H, Takeuchi K, et al. Treatment of lung cancer with an ALK inhibitor after EML4-ALK fusion gene detection using endobronchial ultrasound-guided transbronchial needle aspiration. J Thorac Oncol. 2010;5(12):2041–2043.
    1. Garcia-Olivé I, Monsó E, Andreo F, et al. Endobronchial ultrasound-guided transbronchial needle aspiration for identifying EGFR mutations. Eur Respir J. 2010;35(2):391–395.
    1. Schmid-Bindert G, Wang Y, Jiang H, et al. EBUS-TBNA provides highest RNA yield for multiple biomarker testing from routinely obtained small biopsies in non-small cell lung cancer patients – a comparative study of three different minimal invasive sampling methods. PLoS One. 2013;8(10):e77948.
    1. Kojima K, April C, Canasto-Chibuque C, et al. Transcriptome profiling of archived sectioned formalin-fixed paraffin-embedded (AS-FFPE) tissue for disease classification. PLoS One. 2014;9(1):e86961.
    1. Fedorowicz G, Guerrero S, Wu TD, Modrusan Z. Microarray analysis of RNA extracted from formalin-fixed, paraffin-embedded and matched fresh-frozen ovarian adenocarcinomas. BMC Med Genomics. 2009;2:23.
    1. Freidin MB, Bhudia N, Lim E, Nicholson AG, Cookson WO, Moffatt MF. Impact of collection and storage of lung tumor tissue on whole genome expression profiling. J Mol Diagn. 2012;14(2):140–148.
    1. Lee HS, Lee GK, Lee HS, et al. Real-time endobronchial ultrasound-guided transbronchial needle aspiration in mediastinal staging of non-small cell lung cancer: how many aspirations per target lymph node station? Chest. 2008;134(2):368–374.
    1. Kanaji N, Bandoh S, Ishii T, et al. Cytokeratins negatively regulate the invasive potential of lung cancer cell lines. Oncol Rep. 2011;26(4):763–768.

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