PURPOSE: The therapeutic choice for patients with lung adenocarcinoma depends on the presence of EGFR mutations. In many cases, only cytological samples are available for molecular diagnosis. Bronchoalveolar lavage (BAL) and pleural fluid (PF), which represent a considerable proportion of cytological specimens, cannot always be used for molecular testing because of low rate of tumor cells. EXPERIMENTAL DESIGN: We tested the feasibility of EGFR mutation analysis on BAL and PF samples by Next-Generation Sequencing (NGS), an innovative and extremely sensitive platform. The study was devised to extend the EGFR test to those patients who could not get it due to paucity of biological material. A series of 830 lung cytology specimens was used to select 48 samples (BAL and PF) from patients with EGFR mutations in resected tumors. These samples included 36 cases with 0,3-9% of neoplastic cells (series-A) and 12 cases without evidence of tumor (series-B). All samples were analysed by Sanger sequencing (SS) and NGS on 454 Roche platform. A mean of 21,130 -/+ 2,370 sequences per sample were obtained by NGS. RESULTS: In series-A, EGFR mutations were detected in 16% of cases by SS and in 81% of cases by NGS. Seventy-seven% of cases found to be negative by SS showed mutations by NGS. In series-B, all samples were negative for EGFR mutation by SS, whereas 42% of them were positive by NGS. CONCLUSIONS: The very sensitive EGFR-NGS-assay may open up to the possibility of specific treatments for patients otherwise doomed to re-biopsies or non-targeted therapies.

Effective assessment of egfr mutation status in bronchoalveolar lavage and pleural fluids by next-generation sequencing.

BUTTITTA, Fiamma;FELICIONI, LAURA;DEL GRAMMASTRO, MAELA;FILICE, GIAMPAOLO;MALATESTA, SARA;VIOLA, Patrizia;ZAPPACOSTA, ROBERTA;ROSINI, Sandra;CUCCURULLO, Franco;MARCHETTI, Antonio
2013

Abstract

PURPOSE: The therapeutic choice for patients with lung adenocarcinoma depends on the presence of EGFR mutations. In many cases, only cytological samples are available for molecular diagnosis. Bronchoalveolar lavage (BAL) and pleural fluid (PF), which represent a considerable proportion of cytological specimens, cannot always be used for molecular testing because of low rate of tumor cells. EXPERIMENTAL DESIGN: We tested the feasibility of EGFR mutation analysis on BAL and PF samples by Next-Generation Sequencing (NGS), an innovative and extremely sensitive platform. The study was devised to extend the EGFR test to those patients who could not get it due to paucity of biological material. A series of 830 lung cytology specimens was used to select 48 samples (BAL and PF) from patients with EGFR mutations in resected tumors. These samples included 36 cases with 0,3-9% of neoplastic cells (series-A) and 12 cases without evidence of tumor (series-B). All samples were analysed by Sanger sequencing (SS) and NGS on 454 Roche platform. A mean of 21,130 -/+ 2,370 sequences per sample were obtained by NGS. RESULTS: In series-A, EGFR mutations were detected in 16% of cases by SS and in 81% of cases by NGS. Seventy-seven% of cases found to be negative by SS showed mutations by NGS. In series-B, all samples were negative for EGFR mutation by SS, whereas 42% of them were positive by NGS. CONCLUSIONS: The very sensitive EGFR-NGS-assay may open up to the possibility of specific treatments for patients otherwise doomed to re-biopsies or non-targeted therapies.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11564/383283
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