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A. Estival



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    MO16 - Prognostic and Predictive Biomarkers IV (ID 97)

    • Event: WCLC 2013
    • Type: Mini Oral Abstract Session
    • Track: Medical Oncology
    • Presentations: 1
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      MO16.08 - Cytology samples (s) for EGFR, KRAS and ALK testing in Non-Small-Cell Lung Cancer (NSCLC) (ID 2439)

      16:15 - 17:45  |  Author(s): A. Estival

      • Abstract
      • Presentation
      • Slides

      Background
      Recent advances in targeted therapy in NSCLC have achieved impressive results in advanced disease. For molecular testing,cytology samples are not commonly used since is less likely to be adequate. At ICO Badalona- Germans Trias i Pujol Hospital we have used cytology specimens when biopsy was not available. We describe the general results when using cytology specimens in NSCLC to detect EGFR mutation, KRAS mutations and ALK translocations.

      Methods
      From February 2007 to May 2012, 227 cytology samples from patients with NSCLC were collected at the Department of Pathology as cell block or fresh specimen over an apropiate slide. After that, tumor cells were(8-150) captured by laser microdissection. DNA sequencing for EGFR exons 18, 19, 20, 21, KRAS codons 12 and 13 was performed at Molecular Biology Laboratory( ICO-Badalona) and ALK translocation were analyzed at Pathology Department by FISH

      Results
      EGFR mutations were tested in 227 samples.The overall output was 86.3% (not evaluable in 15 , insufficient tissue in 8, no tumor cells in 4, not done in 4). EGFR mutation was detected in 8.81% (20/227). KRAS mutation were tested in 41 samples with results in 33, 80.5% (2 not evaluable, insufficient tumor cells 3, no tumor 1 and not done 2 samples). KRAS mutation was positive 6 (14.6%). ALk translocation were tested in 9 p with results in 6 p ( 1 not evaluable and 2 insufficient tumor cells) Both cell-block and fresh specimen over an apropiate slide were used to perform molecular testing. The output for cell-block was 83.3%(124/148) and testing was not possible in 23(11 not evaluable, 6 insufficient tumor cells, 4 not tumor and 3 not done). The output for membrane was 91.1% (72/79) and was not possible in 7(4 were not evaluable, 2 insufficient tumor cells and not done in 1). 54.7% of samples were obtained from endobronquial ultrasound guided transbronquial needle aspiration of mediastinal adenopathies, 11.3% lung mass needle aspiration and 11.7% from pleural effusion.

      Conclusion
      Our results support the potential use of cytology samples for molecular testing in NSCLC when biopsy specimens are not available. Both membrane preparations and cytology blocks have been used and are equally suitable for molecular testing.

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    P2.10 - Poster Session 2 - Chemotherapy (ID 207)

    • Event: WCLC 2013
    • Type: Poster Session
    • Track: Medical Oncology
    • Presentations: 1
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      P2.10-035 - EGFR mutation (m) status in serum only to treat Non Small Cell Lung Cancer (NSCLC) patients (p) (ID 2412)

      09:30 - 16:30  |  Author(s): A. Estival

      • Abstract

      Background
      Treatment of EGFR mutated NSCLC p with EGFR TKIs in phase III trials has shown improved efficacy to standard chemotherapy.However, it can be difficult to obtain sufficient tumor tissue for analysis of EGFR m status in a large proportion of p. Nevertheless,so far,no data exists for NSCLC p treated according to EGFR m status in serum alone.

      Methods
      We reviewed our database to identify EGFR mutated p, excluding those for whom status was available in both serum and tissue.We analyzed p treated with an EGFR TKI for whom EGFR mutation status was known in serum only(status in tissue unknown due to insufficient material).At the same time, we reviewed p in whom EGFR m status in tissue was available over the same period in order to compare clinical characteristics and efficacy parameters,PFS,ORR and Overall Survival(OS). EGFR m analysis was performed in cell free circulating DNA(cfDNA)isolated from serum and plasma using the QIAmp DNA blood mini kit.

      Results
      9 p with EGFR m detected in serum and 33 p with EGFR m in tissue were included. In EGFR mutated p in serum, median age 63; male 55.6%; non-smokers 33.3%; former smokers 44.4%; ECOG PS 0-1 66.7%; adenocarcinoma 77.8%; deletion19 33.3%, L858R 66.7%; EGFR TKI treatment in 1st line 44.4%; 2nd or 3rd line 55.6%. ORR: complete response (CR) 22.2%; partial response (PR) 22.2%; stable disease (SD) 22.2%; progressive disease (PD) 11.1%. 2p had poor PS and died prior to evaluation. mPFS 4.7 months (mo). mOS 18 mo. In p with EGFR m in tissue, median age 61; male 36.4%; non-smokers 75.8%; former smokers 24.2%; adenocarcinoma 87.9%; deletion19 75.8%; L858R 24.2%; 1st line 54.5%; 2nd or 3rd line 45.5%. ORR: CR 15.2%; PR 57.6%; SD 12.1%; PD 15.2%. mPFS 8.9 mo. mOS 32.7 mo. The multivariate analysis for OS considering EGFR m in serum differed according to PS (PS 0-1 16.6 mo vs PS > 2 5.2 mo)

      Conclusion
      Obtaining sufficient tissue from NSCLC p for analysis of EGFR m status and other molecular alterations can be difficult. Determination of EGFR m in serum alone is feasible, yields similar results to mutation status in tissue, and could permit us to take treatment decisions.

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    P2.18 - Poster Session 2 - Pathology (ID 176)

    • Event: WCLC 2013
    • Type: Poster Session
    • Track: Pathology
    • Presentations: 1
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      P2.18-006 - HER2 and BRAF mutations in non-small-cell lung cancer (NSCLC) patients (p). (ID 1243)

      09:30 - 16:30  |  Author(s): A. Estival

      • Abstract

      Background
      Treatment of non-small-cell lung cancer (NSCLC) has changed drastically in recent years with an increase in rates of detection of driver mutations in a subgroup of NSCLC patients (p); the most recent mutations described in NSCLC are HER2 and BRAF. Overexpression of HER-2 (insertion in exon 20) occurs in 2-6% of NSCLC patients and is more common in women, never-smokers, Asian and adenocarcinoma hystology (adc). BRAF is found in 1.6-4.9% of p, with a higher rate of V600E mutations in smokers, and non-V600E mutations in non-smokers.

      Methods
      We systematically analyzed HER2 and BRAF mutations in EGFR wild type and non-translocated ALK p and retrospectively reviewed the results. Forty six NSCLC p were included between December 2011 to June 2013. Clinical characteristics such as histological subtype, sex, age, smoking status, and mutational status were analyzed. BRAF mutation was assessed by Taqman based assay (5’ nuclease activity) in an AB 7900HT system, with specific primers and probes for V600E positions. HER2 insertion was assessed using Sanger sequencing. All samples were PCR amplified and sequenced in AB 3130, using specific primers for exon 20.

      Results
      All patients were Caucasian; 25 p (54.3%) were women and 21 p (45.7%) men. Fourteen p (30.4%) were never-smokers, 11 (23.9%) former smokers and 19 (41.3%) smokers. Median age was 58.74 years. Forty one p (89.1%) had adc, 2 had squamous cell histology and 3 had another histology (1carcinod tumour, 1 large cell lung cancer and 1 poorly differentiated NSCLC) . Two p (4.3%) had BRAF V600E mutation: 1 female and 1 male, with median age of 56.5, both smokers. One 65 year old, female, never-smoker p (2.2%) had HER-2 insertion. All mutations were found in adc.

      Conclusion
      Analysis of less common driver mutations such as BRAF and Her2 mutations is feasible for daily clinical practice and could be useful to decide treatment. In our European population, incidence of BRAF and HER-2 mutation is similar to that previously reported (4.3% and 2.2%). Both p with BRAF V600E mutation were smokers, and the p with mutation in HER-2 was a 65 year-old female never-smoker.