The feasibility of molecular testing on cell blocks created from brush tip washings in the assessment of peripheral lung lesions

Bonney, Asha; Christie, Michael; Beaty, Anne; Lunke, Sebastian; Taylor, Graham P.; Irving, Louis; Steinfort, Daniel P.

doi:10.21037/jtd.2016.08.85

Cited by 14 publications

(14 citation statements)

References 16 publications

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“…Nevertheless, the diagnosis of peripheral pulmonary lesions (PPLs) is still a challenge in clinical practice because of the difficulty of approaching them with standard bronchoscopy and because of the need to obtain large samples for pathological and molecular diagnoses . R‐EBUS–guided transbronchial biopsy using fluoroscopy has proven to be the most effective noninvasive techniques for the pathological diagnosis of peripheral lung nodules, and the molecular profiling of these samples seems to be feasible when biopsy or cell blocks can be performed …”

Section: Introductionmentioning

confidence: 99%

Molecular analysis of peripheral lung adenocarcinoma in brush cytology obtained by EBUS plus fluoroscopy‐guided bronchoscopy

Sánchez-Font

Chalela

Martín-Ontiyuelo

et al. 2018

Cancer Cytopathology

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BACKGROUND: More than 60% of patients with lung cancer are diagnosesd at advanced stages. The introduction of targeted therapies requires molecular diagnosis to guide treatment. The aim of this study was to evaluate the feasibility of performing mutational analysis with brushing specimens obtained by radial-miniprobe endobronchial ultrasound (R-EBUS) plus fluoroscopy-guided bronchoscopy in patients with peripheral pulmonary adenocarcinoma. METHODS:Brushing specimens were deposited on cytological slides and were conserved in Roswell Park Memorial Institute (RPMI) culture medium. DNA was isolated to perform a mutational analysis with real-time quantitative polymerase chain reaction and Sanger sequencing for epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS). RESULTS: Thirty patients with adenocarcinoma were prospectively included. In 100% of the patients, the molecular study was viable with brushing specimens. In 16 (53.3%), KRAS or EGFR mutations were detected: 10 KRAS mutations (33.3%) and 6 EGFR mutations (20%). In a comparison with histological samples, a correlation of 86.6% was detected, and only 2 patients with wild-type results from brushing specimens presented with an EGFR mutation in histological samples. CONCLUSIONS: Brushing specimens conserved in RPMI medium and obtained by R-EBUS plus fluoroscopy-guided bronchoscopy are valid for molecular studies. They allow the detection of EGFR/KRAS mutations in patients with peripheral adenocarcinoma. In addition, invasive techniques are avoided, the risk of complications is reduced, and the obtained samples are optimized. Cancer Cytopathol 2018;126:860-871.

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Section: Introductionmentioning

confidence: 99%

Molecular analysis of peripheral lung adenocarcinoma in brush cytology obtained by EBUS plus fluoroscopy‐guided bronchoscopy

Sánchez-Font

Chalela

Martín-Ontiyuelo

et al. 2018

Cancer Cytopathology

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“…EBUS-transbronchial needle aspiration, 4 EBUSguided fine needle aspirates, 5 and bronchoscope cytology brush tip washings can provide adequate material for both immunohistochemical studies 6 and mutation sequencing of EGFR, KRAS, and BRAF genes. 7 Demand is increasing on molecular and sequencing technology to adapt with limited amounts of fixed tumor tissue, which are becoming the most commonly used samples for diagnostic and molecular pathology. However, fixed tumor tissue may not be optimal in all settings or for all downstream analyses.…”

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confidence: 99%

A Novel Approach to Detect Programed Death Ligand 1 (PD-L1) Status and Multiple Tumor Mutations Using a Single Non–Small-Cell Lung Cancer (NSCLC) Bronchoscopy Specimen

Vannitamby

Hendry

Makadia

et al. 2019

The Journal of Molecular Diagnostics

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CME Accreditation Statement: This activity ("JMD 2019 CME Program in Molecular Diagnostics") has been planned and implemented in accordance with the accreditation requirements and policies of the Accreditation Council for Continuing Medical Education (ACCME) through the joint providership of the American Society for Clinical Pathology (ASCP) and the American Society for Investigative Pathology (ASIP). ASCP is accredited by the ACCME to provide continuing medical education for physicians. The ASCP designates this journal-based CME activity ("JMD 2019 CME Program in Molecular Diagnostics") for a maximum of 18.0 AMA PRA Category 1 Credit(s) ä . Physicians should claim only credit commensurate with the extent of their participation in the activity.

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“…Use of highly sensitive molecular techniques may also be of diagnostic utility in other lung cancer specimen types, such as bronchoscopic specimens (19). Such specimens may be non-diagnostic or not suitable for molecular testing (20,21), due to small specimen volume, or due to presence of a significant proportion of non-malignant cells (22). Molecular testing even on a single bronchial brushing specimen may significantly enhance diagnostic sensitivity of minimally invasive investigation (23,24), though this remains to be examined in future studies.…”

Section: Discussionmentioning

confidence: 99%

Sensitive molecular testing methods can demonstrate NSCLC driver mutations in malignant pleural effusion despite non-malignant cytology

Steinfort

Kranz

Dowers

et al. 2019

Transl. Lung Cancer Res.

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Malignant pleural effusion (MPE) may be diagnosed by cytologic evaluation of pleural fluid, though false negative results can occur. Pleural effusions may provide a source of tumour material for genotyping in lung cancer patients. Detection of MPE may be improved through use of highly sensitive molecular techniques. We identified five patients with non-small cell lung cancer (NSCLC) with initial pleural fluid samples that were non-malignant on cytology, but were subsequently clinically confirmed to have MPE. Tumour mutation status was confirmed via routine testing of diagnostic clinical specimens. Cytologically negative pleural fluid cell-block specimens were analysed by amplicon-based parallel sequencing (APS) for somatic mutations commonly detected in NSCLC, and selected cases by improved and complete enrichment CO-amplification at lower denaturation temperature PCR (ICECOLD PCR) for known mutations. Mutations were detected in three out of three (sensitivity 100%) cytologically non-malignant pleural fluids from patients with a known mutation: two patients with known Kirsten rat sarcoma (KRAS) mutation demonstrated the same KRAS mutation in their pleural fluids by APS, both at approximately 2% mutant allele frequency. In one patient with a known KRAS mutation, ICECOLD PCR detected the same KRAS variant at 0.7% frequency. No mutations were detected in patients with wild-type findings from reference samples (specificity 100%). Sensitive DNA sequencing methods can detect cancerdriver mutations in cytologically non-malignant pleural fluid specimens from NSCLC patients with MPE. Our findings demonstrate the feasibility of sensitive molecular diagnostic techniques for improvement of diagnostic assessment of pleural effusions in patients with lung cancer.

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The feasibility of molecular testing on cell blocks created from brush tip washings in the assessment of peripheral lung lesions

Abstract: The findings of this study demonstrate that molecular testing can be performed on cell blocks created from BTW and this technique may allow for an increase in bronchoscope specimens amenable to molecular testing without further increasing morbidity and mortality.

Cited by 14 publications

References 16 publications

Molecular analysis of peripheral lung adenocarcinoma in brush cytology obtained by EBUS plus fluoroscopy‐guided bronchoscopy

Molecular analysis of peripheral lung adenocarcinoma in brush cytology obtained by EBUS plus fluoroscopy‐guided bronchoscopy

A Novel Approach to Detect Programed Death Ligand 1 (PD-L1) Status and Multiple Tumor Mutations Using a Single Non–Small-Cell Lung Cancer (NSCLC) Bronchoscopy Specimen

Sensitive molecular testing methods can demonstrate NSCLC driver mutations in malignant pleural effusion despite non-malignant cytology

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