Pyrosequencing Analysis of EGFR and KRAS Mutations in EUS and EBUS-Derived Cytologic Samples of Adenocarcinomas of the Lung

Stigt, Jos A.; Hart, Nils A 't; Knol, Ageeth J.; Uil, Steven M.; Groen, Harry J.M.

doi:10.1097/jto.0b013e31829ce93e

Cited by 29 publications

(42 citation statements)

References 33 publications

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“…Several groups have shown the retrospective analysis of fixed EBUS-TBNA samples can be used to identify clinically actionable mutations in lung cancer [29]–[33], and that high quality DNA, RNA and protein can be obtained from these samples [34], [35]. Our data extend these studies by demonstrating that freshly isolated EBUS-TBNA samples can generate high-quality DNA suitable for massively parallel targeted re-sequencing that avoids formalin artefact, and that can be carefully controlled for stromal artefact.…”

Section: Discussionmentioning

confidence: 99%

Genomic Characterisation of Small Cell Lung Cancer Patient-Derived Xenografts Generated from Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Specimens

et al. 2014

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Patient-derived xenograft (PDX) models generated from surgical specimens are gaining popularity as preclinical models of cancer. However, establishment of PDX lines from small cell lung cancer (SCLC) patients is difficult due to very limited amount of available biopsy material. We asked whether SCLC cells obtained from endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) could generate PDX lines that maintained the phenotypic and genetic characteristics of the primary tumor. Following successful EBUS-TBNA sampling for diagnostic purposes, we obtained an extra sample for cytologic analysis and implantation into the flanks of immunodeficient mice. Animals were monitored for engraftment for up to 6 months. Histopathologic and immunohistochemical analysis, and targeted next-generation re-sequencing, were then performed in both the primary sample and the derivative PDX line. A total of 12 patients were enrolled in the study. EBUS-TBNA aspirates yielded large numbers of viable tumor cells sufficient to inject between 18,750 and 1,487,000 cells per flank, and to yield microgram quantities of high-quality DNA. Of these, samples from 10 patients generated xenografts (engraftment rate 83%) with a mean latency of 104 days (range 63–188). All but one maintained a typical SCLC phenotype that closely matched the original sample. Identical mutations that are characteristic of SCLC were identified in both the primary sample and xenograft line. EBUS-TBNA has the potential to be a powerful tool in the development of new targeting strategies for SCLC patients by providing large numbers of viable tumor cells suitable for both xenografting and complex genomic analysis.

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Section: Discussionmentioning

confidence: 99%

Genomic Characterisation of Small Cell Lung Cancer Patient-Derived Xenografts Generated from Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Specimens

et al. 2014

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“…68 Cell blocks have been shown to be comparable to core biopsies in some studies for detection of clinically important mutations. 3,69 A significant number of studies have shown that smears and CBs are equivalent for molecular testing. 66,70–72 In an analysis of 181 articles focusing on EGFR analysis in lung cancer, da Cunha Santos et al 64 showed that cytologic techniques using an array of fixation and processing techniques yielded mutation detection rates similar if not superior to histologic material.…”

Section: Committee IVmentioning

confidence: 99%

“…3,65–69 While FFPE material is one of the most common sources of material for molecular analysis in many laboratories, 67, 72 recent studies have shown that material obtained from air-dried and alcohol-fixed cytologic smears can be reliably used. 61,62,73,74 Sample adequacy for analysis should be confirmed by a pathologist.…”

Section: Committee IVmentioning

confidence: 99%

Utilization of ancillary studies in the cytologic diagnosis of respiratory lesions: The papanicolaou society of cytopathology consensus recommendations for respiratory cytology

Layfield

Roy‐Chowdhuri

Baloch

et al. 2016

Diagnostic Cytopathology

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The Papanicolaou Society of Cytopathology has developed a set of guidelines for respiratory cytology including indications for sputum examination, bronchial washings and brushings, CT-guided FNA and endobronchial ultrasound guided fine needle aspiration (EBUS-FNA), as well as recommendations for classification and criteria, ancillary testing and post-cytologic diagnosis management and follow-up. All recommendation documents are based on the expertise of committee members, an extensive literature review, and feedback from presentations at national and international conferences. The guideline documents selectively present the results of these discussions. The present document summarizes recommendations for ancillary testing of cytologic samples. Ancillary testing including microbiologic, immunocytochemical, flow cytometric, and molecular testing, including next-generation sequencing are discussed.

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“…Of the remaining 140 patients, 85% had sufficient material for EGFR and kras testing (12 had no DNA, 4 had unsuccessful amplifications, and 5 had insufficient DNA). 40 …”

Section: Endobronchial Ultrasound Bronchoscopymentioning

confidence: 97%

Tissue Sample Adequacy for Mutational Analysis by Endbronchial Ultrasound and Electromagnetic Navigation Bronchoscopy

Sridhar

Walker

Bowling

et al. 2014

Clinical Pulmonary Medicine

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Lung cancer is the leading cause of cancer death requiring more efficient diagnostic tools for proper and early diagnosis of lung cancer. Common mutations are tested for in adenocarcinoma and squamous cell carcinoma, with the most common ones being epidermal growth factor receptor and thyroid transcription factor-1 for adenocarcinoma. Testing for these mutations is important in planning the proper course of treatment. Previously mediastinoscopy, was the gold standard to extract adequate samples for molecular testing, however, this procedure has a low sensitivity and complications associated with it. Newer approaches to diagnosis of lung cancer include endobronchial ultrasound transbronchial needle aspiration and electromagnetic navigation bronchoscopy. Endobronchial ultrasound is a real-time technique that allows for visualization of the sample and the needle using an ultrasound. From commuted tomography scans taken of the chest, electromagnetic navigation bronchoscopy constructs a 3-dimensional image of the lungs with a highlighted navigation to the tumor. In reviewing numerous papers, it was seen that the use of these techniques for sample collection provides a high percentage of sample adequacy with increased sensitivity and specificity. The use of these techniques is becoming invaluable in properly diagnosing and staging lung cancers with minimal complications.

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Pyrosequencing Analysis of EGFR and KRAS Mutations in EUS and EBUS-Derived Cytologic Samples of Adenocarcinomas of the Lung

Abstract: Pyrosequencing on cytological blocks, especially older tumor blocks, is feasible with a high diagnostic success rate. Failures in HRM were observed in DNA samples with short fragments related to longer storage times.

Cited by 29 publications

References 33 publications

Genomic Characterisation of Small Cell Lung Cancer Patient-Derived Xenografts Generated from Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Specimens

Genomic Characterisation of Small Cell Lung Cancer Patient-Derived Xenografts Generated from Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Specimens

Utilization of ancillary studies in the cytologic diagnosis of respiratory lesions: The papanicolaou society of cytopathology consensus recommendations for respiratory cytology

Tissue Sample Adequacy for Mutational Analysis by Endbronchial Ultrasound and Electromagnetic Navigation Bronchoscopy

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