Multigene Mutation Analysis of Metastatic Lymph Nodes in Non-small Cell Lung Cancer Diagnosed by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration

doi:10.1378/chest.10-3186

Chest

Volume 140, Issue 5, November 2011, Pages 1319-1324

https://doi.org/10.1378/chest.10-3186 Get rights and content

Background

The importance of biomarker analysis in patients with non-small cell lung cancer (NSCLC) is well known. The purpose of this study was to analyze the mutation status of multiple genes in metastatic lymph nodes obtained by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) and to examine the correlation between treatments and outcomes.

Methods

Genetic alterations were analyzed in metastatic hilar or mediastinal lymph nodes diagnosed by EBUS-TBNA in 156 patients with NSCLC. Epidermal growth factor receptor (EGFR) was analyzed using the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method (n = 156). V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-ras) (exons 2-3) and tumor protein 53 (p53) (exons 4-8) were analyzed by direct sequencing (n = 113). In addition, retrospective chart review was performed for clinical data analysis.

Results

EGFR gene mutations were detected in 42 cases (26.9%). Twenty-three patients with EGFR mutations received gefitinib, with an overall response rate (partial response [PR]) of 54.5% and disease control rate (PR + stable disease) of 86.4% (Response Evaluation Criteria in Solid Tumors). K-ras gene mutations were detected in four cases (3.5%), and p53 gene mutations were detected in 47 cases (41.6%). Fifty-two patients underwent conventional chemotherapy (46 patients underwent platinum-based chemotherapy). Patients with p53 gene mutations showed chemoresistance (progressive disease of 42.9%, P = .0339) and a relatively poor prognosis after chemotherapy (P = .1391).

Conclusions

Multigene mutation analysis can be performed in EBUS-TBNA samples of metastatic lymph nodes from patients with NSCLC. EBUS-TBNA allows genetic evaluation of tumor cells within the metastatic node, which may allow physicians to better select treatments, particularly EGFR tyrosine kinase inhibitors.

Section snippets

Patients

From April 2008 to December 2009, 156 consecutive NSCLC cases with proven hilar and/or mediastinal lymph node metastasis diagnosed by EBUS-TBNA were enrolled. An independent pathologist reviewed all cases and pathologically confirmed the diagnosis of NSCLC in each specimen. EGFR gene mutation assessment was routinely performed in EBUS-TBNA samples when pathology confirmed metastatic disease in the lymph node. A retrospective chart review was performed for all patients. Response to chemotherapy

Patient Characteristics

The clinical characteristics of the 156 patients are listed in Table 1. All cases had hilar and/or mediastinal lymph node metastasis confirmed by EBUS-TBNA. Overall, 138 mediastinal and 18 hilar lymph nodes were evaluated. The median lymph node size was 14.0 mm (range, 4.8-33.4 mm) in the short axis on ultrasound image.

EGFR Gene Mutation Assessment

Mediastinal lymph nodes were first used for EGFR mutation analysis followed by hilar lymph nodes if necessary. EGFR mutation analysis was possible in 154 out of 156 cases (98.7%)

Discussion

We have previously reported the usefulness of EBUS-TBNA for EGFR gene mutation assessment as well as pathologic diagnosis.⁵ EBUS-TBNA is a minimally invasive modality that is safe and has a high diagnostic yield for tissue diagnosis of mediastinal and hilar lymphadenopathy in patients with NSCLC.9, 10, 11 Moreover, biopsy samples obtained with EBUS-TBNA can be subjected to molecular analysis in addition to pathologic evaluation, the clinical significance of which is increasing as molecularly

Acknowledgments

Author contributions: Dr Nakajima: contributed to study design, development of methodology, collection of data, analysis and interpretation of data, wrote all sections of the manuscript, and read and approved the final version of this manuscript.

Dr Yasufuku: contributed to analysis and interpretation of data, supervision, and read and approved the final version of this manuscript.

Dr Nakagawara: contributed to study design and supervision and read and approved the final version of this

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Next-Generation Sequencing for Genotyping of Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Samples in Lung Cancer
2019, Annals of Thoracic Surgery
Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) can obtain a small amount of specimen. This study aims to evaluate the feasibility and robustness of using EBUS-EBNA samples to perform capture-based targeted next-generation sequencing (NGS).
Tissue samples from patients with advanced non-small cell lung cancer were collected by EBUS-TBNA and were formalin-fixed paraffin-embedded. Three representative genes, EGFR, ALK, and ROS1, were examined by amplification refractory mutation system polymerase chain reaction, immunohistochemistry, and quantitative reverse transcription polymerase chain reaction. The remaining samples were processed with NGS assay with a 56-gene panel. Classic driver mutations detected by NGS were verified by conventional methods.
Of the 85 samples from patients with advanced non-small cell lung cancer, 77 were performed successfully with all assays. Forty-one mutations in EGFR, ALK, and ROS1 were detected in both conventional methods and NGS, representing a 100% concordance. In contrast, four EGFR mutations detected by NGS were not covered in the targeted regions of amplification refractory mutation system polymerase chain reaction, leading to a negative call in these patients. Altogether, NGS detected 12 additional variants, including six KRAS mutations, one BRAF mutation, one RET fusion, one MET amplification concurrent with EGFR L858R, one KRAS amplification together with EGFR 19del, and one ERBB2 amplification. The mean number of needle passes per lymph node was 5.2 in samples successfully applied in all assays.
NGS assay can be successfully conducted with limited tissue samples obtained from EBUS-TBNA. Compared with conventional methods, NGS assay provides more comprehensive information on genetic alterations in tumors, which greatly assists therapeutic decision making for advanced lung cancer.
Pan-asian adapted clinical practice guidelines for the management of patients with metastatic non-small-cell lung cancer: A csco-esmo initiative endorsed by jsmo, ksmo, mos, sso and tos
2019, Annals of Oncology
The most recent version of the European Society for Medical Oncology (ESMO) Clinical Practice Guidelines for the diagnosis, treatment and follow-up of metastatic non-small-cell lung cancer (NSCLC) was published in 2016. At the ESMO Asia Meeting in November 2017 it was decided by both ESMO and the Chinese Society of Clinical Oncology (CSCO) to convene a special guidelines meeting immediately after the Chinese Thoracic Oncology Group Annual Meeting 2018, in Guangzhou, China. The aim was to adapt the ESMO 2016 guidelines to take into account the ethnic differences associated with the treatment of metastatic NSCLC cancer in Asian patients. These guidelines represent the consensus opinions reached by experts in the treatment of patients with metastatic NSCLC representing the oncological societies of China (CSCO), Japan (JSMO), Korea (KSMO), Malaysia (MOS), Singapore (SSO) and Taiwan (TOS). The voting was based on scientific evidence, and was independent of both the current treatment practices and the drug availability and reimbursement situations in the six participating Asian countries. During the review process, the updated ESMO 2018 Clinical Practice Guidelines for metastatic NSCLC were released and were also considered, during the final stages of the development of the Pan-Asian adapted Clinical Practice Guidelines.
Metastatic non-small cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up
2018, Annals of Oncology
Comparison of Programmed Death Ligand-1 Immunohistochemical Staining Between Endobronchial Ultrasound Transbronchial Needle Aspiration and Resected Lung Cancer Specimens
2018, Chest
In advanced non-small cell lung cancer (NSCLC), small biopsy specimens from endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) are often the only available material from cancer tissue for the analysis of programmed death ligand-1 (PD-L1) expression. We aim to assess the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of PD-L1 expression at ≥ 1% and ≥ 50% on EBUS-TBNA samples compared with their corresponding surgically resected tumor.
We retrospectively reviewed all patients who underwent EBUS-TBNA followed by surgical resection of NSCLC between July 2006 and September 2016. Demographic information and periprocedural/surgical data were collected. The archived specimens were retrieved and assessed for PD-L1. A positive PD-L1 stain was defined using two separate cutoff points: ≥ 1% and ≥ 50% of tumor cell positivity. EBUS-TBNA aspirates were compared with the surgically resected specimen to calculate the sensitivity, specificity, PPV, and NPV.
Sixty-one patients were included. For PD-L1 ≥ 1%, the sensitivity, specificity, PPV, and NPV were 72%, 100%, 100%, and 80%, respectively. For PD-L1 ≥ 50%, the sensitivity, specificity, PPV, and NPV were 47%, 93%, 70%, and 84%, respectively. The concordance rates for PD-L1 ≥ 1% and ≥ 50% were 87% and 82%, respectively.
A PD-L1 cutoff of ≥ 1% on EBUS-TBNA has a strong correlation with resected tumor specimen. For PD-L1 ≥ 50%, there is a significant decrease in the sensitivity and PPV of EBUS-TBNA specimen when compared with resected tumor. When analyzing for PD-L1 expression using a cutoff of ≥ 50%, EBUS-TBNA specimens may misclassify the status of PD-L1.
Feasibility of Endobronchial Ultrasound-guided Transbronchial Needle Aspiration Cytology Specimens for Next Generation Sequencing in Non–small-cell Lung Cancer
2018, Clinical Lung Cancer
Citation Excerpt :
With these guidelines in mind, endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) has become the procedure of choice to diagnose and stage locally metastatic lung cancer. EBUS-TBNA specimen adequacy for genetic testing varies between 70% and 100% and appears to be equivalent to that from histologic samples.2-5 In these studies, however, testing was for individual genes using direct sequencing, polymerase chain reaction (PCR), or immunohistochemistry, and genetic testing was limited to EGFR, KRAS, BRAF, ALK, and PIK3CA.
Next generation sequencing (NGS) testing of lung cancer is recommended by guidelines, and endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) often provides the only material available for testing. Previous studies have demonstrated successful NGS testing on cell block samples obtained by EBUS; however, cytology smears provide a more reliable sample with better DNA quality for testing. In this study, we aimed to determine the success rate of OncoScreen (50 gene) and OncoPlus (1213 gene) panel NGS testing of cytology samples obtained by EBUS utilizing 22- and 25-gauge needles.
Fifty-four patients underwent EBUS-TBNA of lung cancer for which NGS testing was requested. Data was analyzed for needle gauge, cytologic assessment, NGS test success, and sample type (cytology smear or cell block) used for testing.
Eighty-five NGS tests were ordered on 54 samples. Overall, 95.3% of samples had successful testing. OncoScreen and OncoPlus panels were successful 98.0% and 91.4% of the time, respectively. Cytology smears provided testing material for 85% of the tests. OncoScreen testing was successful in 97.5% and 100% of the 22- and 25-gauge samples, respectively (P = 1.00). OncoPlus testing was successful in 91.3% and 100% of the 22- and 25-gauge samples, respectively (P = 1.00).
NGS can be reliably performed on cytology smears obtained from EBUS-TBNA. The size of the needle does not seem to affect the success rate of small or large panel NGS tests.
Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration for Diagnosis and Staging of Lung Cancer
2018, Clinics in Chest Medicine

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Part of this article has been published in abstract form (Nakajima T, Yasufuku K, Nakagawara A, et al. Chest. 2010;138(4)(suppl 4):728A).

Funding/Support: This research was supported in part by the Ministry of Education, Culture, Sports, Science, and Technology, Grant-in-Aid for Young Scientists (B) [Grant 21791340 in 2009] (T. N.), and Grant-in-Aid for Cancer Research from Ministry of Health, Labor and Welfare in 2009 (T. N.).

Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (http://www.chestpubs.org/site/misc/reprints.xhtml).

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Original ResearchCancerMultigene Mutation Analysis of Metastatic Lymph Nodes in Non-small Cell Lung Cancer Diagnosed by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration

Background

Methods

Results

Conclusions

Section snippets

Patients

Patient Characteristics

EGFR Gene Mutation Assessment

Discussion

Acknowledgments

Lancet Oncol

Chest

Lung Cancer

Chest

Eur J Cancer

Eur J Cancer

Am J Pathol

J Mol Diagn

Chest

J Thorac Oncol

Ann Oncol

J Thorac Oncol

J Thorac Oncol

Original Research
Cancer
Multigene Mutation Analysis of Metastatic Lymph Nodes in Non-small Cell Lung Cancer Diagnosed by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration