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F. Lopez-Rios



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    P1.06 - Poster Session 1 - Prognostic and Predictive Biomarkers (ID 161)

    • Event: WCLC 2013
    • Type: Poster Session
    • Track: Biology
    • Presentations: 1
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      P1.06-023 - Anaplastic Lymphoma Kinase (ALK)-detection in Non-small Cell Lung Cancer: results of the first European IHC-based (D5F3-Optiview) panel test within 16 institutes (ID 1825)

      09:30 - 09:30  |  Author(s): F. Lopez-Rios

      • Abstract

      Background
      The study was supported by Ventana Medical Systems, Inc., a Member of the Roche Group Background: The reliable identification of NSCLC patients with anaplastic lymphoma kinase (ALK) gene rearrangement is crucial for the prescription of ALK tyrosine kinase inhibitors (e.g. crizotinib). Whereas the US FDA-approval (2011) is based upon FISH-testing, the European EMA-approval (2012) refers to the definition of “ALK-positive” NSCLCs without mandating a particular test. Therefore a reliable ALK-immunohistochemistry (IHC) could be a promising option in daily routine practice.

      Methods
      Material and methods: To test the reliability of ALK-IHC-diagnosis in a multi-centre environment (17 European institutes from Belgium, Denmark, France, Germany, Scotland, Spain, Sweden and Switzerland) two tissue microarrays (TMA) consisting of 15 NSCLC cases (all adenocarcinomas; 3 cores for each case) were independently tested for ALK-expression by each laboratory using Ventana Medical System’s ALK (D5F3) primary antibody combined with OptiView DAB IHC detection and OptiView Amplification kits. Cases included in the study were unequivocal ALK-break positive or negative (by FISH), as well as so called “ALK-borderline” cases (low percentage of ALK-break positive cells by FISH, around the cut-off of 15%, therefore challenging in diagnosis, but PCR-confirmed as harbouring EML-4-ALK-fusion variants and thus eligible for therapy). Prior to the TMA-based case testing, each participating instrument was qualified using the VENTANA ALK 2 in 1 Control Slides. To provide a uniform baseline interpretation, a webinar-based training was given to all observers. This training included an overview of the ALK Interpretation Guide, a guided review of 50 patient cases using digital whole slide images, and a proficiency exam certifying each observer.

      Results
      Results: Detailed data analysis was only partly accomplished at the time of abstract submission and will be presented in detail at the “World Conference on LUNG Cancer” in Sydney. Besides the binary evaluation of the cases (ALK-negative vs. ALK-positive) observers were asked to estimate the staining intensity (0-3) within positive cases in correlation to the number of tumor cells and to generate the H-score.

      Conclusion
      Conclusion: Referring to the EMA-approval text our multi-centre study may contribute to validation and accuracy of IHC-based ALK-testing. Such a validated and reliable IHC-assay could be used: (a) as a good pre-screening method reducing time consuming and costly FISH analysis (shorten turn-around time for test results) and (b) as a final predictive approach in cases with reduced interpretability of FISH results (e.g. minimal tumor cell content in small biopsies, decalcified or artificial altered tissue, FISH in doubt/”borderline”).

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    P3.06 - Poster Session 3 - Prognostic and Predictive Biomarkers (ID 178)

    • Event: WCLC 2013
    • Type: Poster Session
    • Track: Biology
    • Presentations: 1
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      P3.06-005 - A Prospective Protocol for Simultaneous Genomic Testing in Patients with Non-Small Cell Lung Cancer (ID 993)

      09:30 - 09:30  |  Author(s): F. Lopez-Rios

      • Abstract

      Background
      Molecular testing in lung cancer has dramatically evolved over the past few years with a large number of targeted therapies and associated biomarkers. This has created a demand for tissue preservation. A careful consensus for prioritization of the different assays is needed to identify the right therapy for the patient. It is necessary to have protocols available that give access to results rapidly and accurately without depleting the sample throughout the process. We have designed a prospective study to demonstrate an efficient biomarker testing workflow (EGFR, ALK and KRAS) in a real clinical setting that preserves limited, valuable clinical samples

      Methods
      This prospective study was conducted at two independent pathology laboratories (Laboratorio de Dianas Terapeuticas of Madrid, Queen’s University of Belfast). Each site performed tissue sectioning for diagnosis and molecular testing according to laboratory protocols or to the assay specific package insert. Digital pathology was used to annotate specimens. NSCLC specimens were obtained by each lab (surgical resections and small biopsies of primary and metastatic lesions), excluding cytology specimens, with the exception of cells blocks. EGFR and KRAS mutation testing was performed using the cobas[®] EGFR Mutation Test and the cobas[®] KRAS Mutation Test (both CE-IVD), using a single 5µm section per test. FISH analysis was performed using the Vysis LSI ALK probe. Figure 1

      Results
      A total of 103 cases were included. 100% of the specimens were successfully tested for EGFR mutation status. Failure rates were 1.9% for ALK and KRAS analysis. Failed DNA extraction/PCR amplification was 3.9%/2.9% for EGFR and 5.8%/12.6% for KRAS. The prevalence of molecular alterations identified in EGFR (4% and 13.2%), ALK (4% and 7.8%) and KRAS (26% and 29.4%) was somewhat similar to that described in earlier studies. ALK rearrangements were mutually exclusive with EGFR and KRAS mutations. The median turnaround time was almost identical between sites: 6-7 days for all biomarkers (EGFR range 1-12 days, ALK range 2-14 days and KRAS range 1-17 days).

      Conclusion
      We have demonstrated that it is feasible to incorporate this protocol into the routine analysis of thoracic samples. The pattern of retesting to achieve full analysis in all samples suggests that, while EGFR testing is easily achievable with a one-section approach, the analysis of KRAS and ALK may require another round of testing in a small percentage of cases. Our results thus provide a pre-analytical and analytical rationale for comprehensive genotyping in patients with NSCLC.