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EGFR T790M mutation testing of non-small cell lung cancer tissue and blood samples artificially spiked with circulating cell-free tumor DNA: results of a round robin trial

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Abstract

The European Commision (EC) recently approved osimertinib for the treatment of adult patients with locally advanced or metastatic non-small-cell lung cancer (NSCLC) harboring EGFR T790M mutations. Besides tissue-based testing, blood samples containing cell-free circulating tumor DNA (ctDNA) can be used to interrogate T790M status. Herein, we describe the conditions and results of a round robin trial (RRT) for T790M mutation testing in NSCLC tissue specimens and peripheral blood samples spiked with cell line DNA mimicking tumor-derived ctDNA. The underlying objectives of this two-staged external quality assessment (EQA) approach were (a) to evaluate the accuracy of T790M mutations testing across multiple centers and (b) to investigate if a liquid biopsy-based testing for T790M mutations in spiked blood samples is feasible in routine diagnostic. Based on a successfully completed internal phase I RRT, an open RRT for EGFR T790M mutation testing in tumor tissue and blood samples was initiated. In total, 48 pathology centers participated in the EQA. Of these, 47 (97.9%) centers submitted their analyses within the pre-defined time frame and 44 (tissue), respectively, 40 (plasma) successfully passed the test. The overall success rates in the RRT phase II were 91.7% (tissue) and 83.3% (blood), respectively. Thirty-eight out of 48 participants (79.2%) successfully passed both parts of the RRT. The RRT for blood-based EGFR testing initiated in Germany is, to the best of our knowledge, the first of his kind in Europe. In summary, our results demonstrate that blood-based genotyping for EGFR resistance mutations can be successfully integrated in routine molecular diagnostics complementing the array of molecular methods already available at pathology centers in Germany.

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Funding

Phase I (internal round robin trial) of this study was funded by AstraZeneca.

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Authors and Affiliations

  1. Institue of Pathology, University Hospital Cologne, Kerpener Strasse 62, 50924, Cologne, Germany

    Jana Fassunke, Michaela Angelika Ihle, Reinhard Büttner & Sabine Merkelbach-Bruse

  2. Institute of Pathology, Charité—University Hospital Berlin, Campus Mitte, Virchowweg 15, 10117, Berlin, Germany

    Dido Lenze, Annika Lehmann, Michael Hummel, Claudia Vollbrecht & Manfred Dietel

  3. Institute of Pathology, University Hospital Heidelberg, Im Neuenheimer Feld 224, Heidelberg, Germany

    Roland Penzel, Anna-Lena Volckmar, Albrecht Stenzinger, Volker Endris & Peter Schirmacher

  4. Pathologisches Institut of the Ludwig-Maximilinas-Universität München, Thalkirchnerstrasse 36, 80336, Munich, Germany

    Andreas Jung & Thomas Kirchner

  5. Institute of Pathology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625, Hannover, Germany

    Ulrich Lehmann & Hans Kreipe

  6. Institute of Pathology, University Hospital Dresden Carl Gustav Carus, Fetscherstrasse 74, 01307, Dresden, Germany

    Silke Zeugner & Gustavo Baretton

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  1. Jana Fassunke
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  2. Michaela Angelika Ihle
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Corresponding author

Correspondence to Jana Fassunke.

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Conflict of interest

Author JF has received speaker honoraria from AstraZeneca and Roche. Author AS has received speaker honoraria from AstraZeneca and is a member of the Diagnostic Content advisory board of AstraZeneca. VE received speaker honoraria from AstraZeneca and is a advisory board members of Astra Zeneca. Author AJ has received speaker honoraria from Amgen, AstraZeneca, Merck-Serono, Roche Pharmaceuticals and is a member of committee (advisory board): Amgen, AstraZeneca, Biocartis, Merck-Serono, Novartis. Author UL has received speaker honoraria from AstraZeneca and Roche. Author HK has received honorarium from Astra Zeneca as an advisory board member. Author PS received speaker honoraria from Astra Zeneca and is an advisory board member of AstraZeneca. Author TK has received speaker honoraria from Merck and AstraZeneca, is a member of AdBoard (Consulting): Amgen, AstraZeneca, Merck KGaA, MSD, Novartis, Pfizer, Roche and has received research grants from Merck and Roche. Author RB has received speaker honoraria from AstraZeneca, Boehringer Ingelheim, BMS, MSD, Pfizer, Qiagen and RocheVentana, has served in Scientific Advisory Boards from AstraZeneca, Boehringer Ingelheim, BMS, MSD, Pfizer, Qiagen and RocheVentana, and is a Cofounder and Chief Scientific Officer of Targos Molecular Pathology, Kassel, Germany. Author SMB has received speaker honoraria from AstraZeneca, Bristol Myers Squibb, Roche and Novartis and has received research grants from Astra Zeneca and Novartis. MI, DL, MH, CV, AL, RP, AV, SZ, GB, and MD declare that they have no conflict of interest.

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The samples were collected with informed consent and under approved local ethical protocols from each patient.

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Fassunke, J., Ihle, M.A., Lenze, D. et al. EGFR T790M mutation testing of non-small cell lung cancer tissue and blood samples artificially spiked with circulating cell-free tumor DNA: results of a round robin trial. Virchows Arch 471, 509–520 (2017). https://doi.org/10.1007/s00428-017-2226-8

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  • DOI: https://doi.org/10.1007/s00428-017-2226-8

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