Endobronchial ultrasound-guided transbronchial needle aspiration for identifying EGFR mutations

García-Olivé, Ignasi; Monsó, Eduard; Andreo, Felipe; Sanz-Santos, José; Tarón, Miguel; Molina-Vila, Miguel Ángel; Llatjós, María; Castellà, Eva; Morán, Teresa; Bertrán-Alamillo, Jordi; Mayo‐de‐las‐Casas, Clara; Queralt, Cristina; Rosell, Rafael

doi:10.1183/09031936.00028109

Cited by 121 publications

(89 citation statements)

References 32 publications

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“…The genes tested have included EGFR KRAS, BRAF and PIK3CA in the lung specimens and BRAF, RET, RAS, TRK and PPRg in the thyroid nodules. 6,7,10,23,[30][31][32][33][34][35][36][37][38] Due to the low analytical sensitivity of direct sequencing, ultrasensitive techniques such as high resolution melting, realtime PCR and pyrosequencing have been utilized to improve detection sensitivity. 7 However, assessment of multiple genes in cytological specimens is sometimes not possible, because traditional platforms require large amounts of DNA.…”

Section: Discussionmentioning

confidence: 99%

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

et al. 2014

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Increasing use of fine needle aspiration for oncological diagnosis, while minimally invasive, poses a challenge for molecular testing by traditional sequencing platforms due to high sample requirements. The advent of affordable benchtop next-generation sequencing platforms such as the semiconductor-based Ion Personal Genome Machine (PGM) Sequencer has facilitated multi-gene mutational profiling using only nanograms of DNA. We describe successful next-generation sequencing-based testing of fine needle aspiration cytological specimens in a clinical laboratory setting. We selected 61 tumor specimens, obtained by fine needle aspiration, with known mutational status for clinically relevant genes; of these, 31 specimens yielded sufficient DNA for next-generation sequencing testing. Ten nanograms of DNA from each sample was tested for mutations in the hotspot regions of 46 cancer-related genes using a 318-chip on Ion PGM Sequencer. All tested samples underwent successful targeted sequencing of 46 genes. We showed 100% concordance of results between next-generation sequencing and conventional test platforms for all previously known point mutations that included BRAF, EGFR, KRAS, MET, NRAS, PIK3CA, RET and TP53, deletions of EGFR and wild-type calls. Furthermore, next-generation sequencing detected variants in 19 of the 31 (61%) patient samples that were not detected by traditional platforms, thus increasing the utility of mutation analysis; these variants involved the APC, ATM, CDKN2A, CTNNB1, FGFR2, FLT3, KDR, KIT, KRAS, MLH1, NRAS, PIK3CA, SMAD4, STK11 and TP53 genes. The results of this study show that next-generation sequencing-based mutational profiling can be performed on fine needle aspiration cytological smears and cell blocks. Next-generation sequencing can be performed with only nanograms of DNA and has better sensitivity than traditional sequencing platforms. Use of next-generation sequencing also enhances the power of fine needle aspiration by providing gene mutation results that can direct personalized cancer therapy.

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Section: Discussionmentioning

confidence: 99%

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

et al. 2014

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“…Kwalifikacja do leczenia celowanego w Centrum Onkologii w Bydgoszczy wynosi średnio 9 dni, licząc od momentu skierowania pacjenta na oznaczenie mutacji w genie EGFR, a kończąc na wydaniu wyniku. Kluczowe znaczenie multidyscyplinarnego zespołu zostało już wcześniej opisane w piśmiennictwie [15][16][17]. Autorzy wskazują, że zastosowanie nowoczesnych technik przez torakochirurgów pozwala na uzyskanie bardziej reprezentatywnych fragmentów tkanki nowotworowej, która później będzie poddana badaniom patomorfologicznym i molekularno-genetycznym.…”

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Analysis of EGFR mutation frequency and coexistence of KRAS and EGFR mutations using RT-PCR in lung adenocarcinoma: may a clinical and pathological model of a patient’s qualification for targeted therapy have an impact on time to obtain genetic results?

Lewandowska¹,

Jóźwicki²,

Starzyński³

et al. 2012

kitp

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StreszczenieW Polsce rak płuca jest najczęstszą przyczyną zgonów spowodowanych chorobą nowotworową wśród mężczyzn, a od 2009 r. także wśród kobiet. W niedrobnokomórkowym raku płuca (NDRP) może dojść do nieprawidłowości w metabolizmie genu EGFR, takich jak silna ekspresja białka EGFR, amplifikacje lub mutacje somatyczne w domenie kinazy tyrozynowej (TK). Leczenie chorych z mutacją aktywującą w genie EGFR inhibitorami TK EGFR jest skuteczniejsze niż standardowa chemioterapia, ale kluczem do sukcesu jest prawidłowa kwalifikacja pacjenta, realizowana dzięki współpracy interdyscyplinarnego zespołu torakochirurgów, patomorfologów, genetyków -biologów molekularnych i onkologów. Kwalifikacja pacjentów do badania obejmowała ocenę kliniczną oraz patomorfologiczną. Badanie genetyczne wykonano przy użyciu metody real-time PCR. Do analizy najczęstszych 29 mutacji w genie EGFR i 7 najczęstszych mutacji w genie KRAS użyto odpowiednio sond hydrolizujących i hybrydyzujących. U dziesięciu z 77 pacjentów wykryto mutację aktywującą, w przeważającej większości w eksonach 19 i 21, co stanowi Ocena częstości występowania mutacji w genie EGFR i współwystępowania mutacji EGFR i KRAS metodą reakcji łańcuchowej polimerazy w czasie rzeczywistym u chorych na raka gruczołowego płuca -czy kliniczno--patologiczny model kwalifikacji pacjenta do leczenia celowanego może mieć wpływ na czas uzyskania wyniku genetycznego?Analysis of EGFR mutation frequency and coexistence of KRAS and EGFR mutations using RT-PCR in lung adenocarcinoma: may a clinical and pathological model of a patient's qualification for targeted therapy have an impact on time to obtain genetic results? AbstractIn Poland, lung cancer is the most common cause of cancer deaths among men, and since 2009 in women as well. Nonsmall cell lung carcinoma (NSCLC) can lead to abnormalities in the metabolism of EGFR, such as strong EGFR protein expression, amplification or presence of somatic mutations in the tyrosine kinase domain (TK). Targeted therapy (EGFR TK inhibitors) for patients diagnosed with activating EGFR mutations is more effective than standard chemotherapy, but the key to success is proper classification of the patient, carried out through the cooperation of an interdisciplinary team: thoracic surgeons, pathologists, geneticists/molecular biologists and oncologists. Eligibility for the study included clinical and pathological assessment. Genetic testing was performed using real-time PCR: we analysed 29 most common EGFR mutations (Entrogen) and 9 most common KRAS mutations (TibMolBiol).

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“…Minimally invasive procedures like endobronchial ultrasound-guided transbronchial needle aspiration are gaining popularity in the staging of advanced lung carcinoma patients because of the multitude of information that can be obtained from the aspirated material, including pathologic diagnosis and molecular testing. 14,15 These minimally invasive techniques also allow for serial sampling of a patient's tumor to assess therapeutic response as well as identify additional molecular markers of resistance. With this recent paradigm shift in lung cancer diagnosis and management, cytology has moved away from being a screening modality or ancillary technique and has established itself as an independent diagnostic procedure that plays a predictive role in determining clinical management.…”

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EGFR and KRAS mutation analysis in cytologic samples of lung adenocarcinoma enabled by laser capture microdissection

Roy‐Chowdhuri

Pham

et al. 2012

Modern Pathology

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The discovery of activating mutations in EGFR and KRAS in a subset of lung adenocarcinomas was a major advance in our understanding of lung adenocarcinoma biology, and has led to groundbreaking studies that have demonstrated the efficacy of tyrosine kinase inhibitor therapy. Fine-needle aspirates and other cytologic procedures have become increasingly popular for obtaining diagnostic material in lung carcinomas. However, frequently the small amount of material or sparseness of tumor cells obtained from cytologic preparations limit the number of specialized studies, such as mutation analysis, that can be performed. In this study we used laser capture microdissection to isolate small numbers of tumor cells to assess for EGFR and KRAS mutations from cell block sections of 19 cytology samples from patients with known lung adenocarcinomas. We compared our results with previous molecular assays that had been performed on either surgical or cytology specimens as part of the patient's initial clinical work-up. Not only were we able to detect the identical EGFR or KRAS mutation that was present in the patient's prior molecular assay in every case, but we were also able to consistently detect the mutation from as few as 50 microdissected tumor cells. Furthermore, isolating a more pure population of tumor cells resulted in increased sensitivity of mutation detection as we were able to detect mutations from laser capture microdissection-enriched cases where the tumor load was low and traditional methods of whole slide scraping failed. Therefore, this method can not only significantly increase the number of lung adenocarcinoma patients that can be screened for EGFR and KRAS mutations, but can also facilitate the use of cytologic samples in the newly emerging field of molecular-based personalized therapies.

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Endobronchial ultrasound-guided transbronchial needle aspiration for identifying EGFR mutations

Cited by 121 publications

References 32 publications

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

Analysis of EGFR mutation frequency and coexistence of KRAS and EGFR mutations using RT-PCR in lung adenocarcinoma: may a clinical and pathological model of a patient’s qualification for targeted therapy have an impact on time to obtain genetic results?

EGFR and KRAS mutation analysis in cytologic samples of lung adenocarcinoma enabled by laser capture microdissection

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