Next-generation sequencing in routine brain tumor diagnostics enables an integrated diagnosis and identifies actionable targets View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2016-06

AUTHORS

Felix Sahm, Daniel Schrimpf, David T. W. Jones, Jochen Meyer, Annekathrin Kratz, David Reuss, David Capper, Christian Koelsche, Andrey Korshunov, Benedikt Wiestler, Ivo Buchhalter, Till Milde, Florian Selt, Dominik Sturm, Marcel Kool, Manuela Hummel, Melanie Bewerunge-Hudler, Christian Mawrin, Ulrich Schüller, Christine Jungk, Antje Wick, Olaf Witt, Michael Platten, Christel Herold-Mende, Andreas Unterberg, Stefan M. Pfister, Wolfgang Wick, Andreas von Deimling

ABSTRACT

With the number of prognostic and predictive genetic markers in neuro-oncology steadily growing, the need for comprehensive molecular analysis of neuropathology samples has vastly increased. We therefore developed a customized enrichment/hybrid-capture-based next-generation sequencing (NGS) gene panel comprising the entire coding and selected intronic and promoter regions of 130 genes recurrently altered in brain tumors, allowing for the detection of single nucleotide variations, fusions, and copy number aberrations. Optimization of probe design, library generation and sequencing conditions on 150 samples resulted in a 5-workday routine workflow from the formalin-fixed paraffin-embedded sample to neuropathological report. This protocol was applied to 79 retrospective cases with established molecular aberrations for validation and 71 prospective cases for discovery of potential therapeutic targets. Concordance of NGS compared to established, single biomarker methods was 98.0 %, with discrepancies resulting from one case where a TERT promoter mutation was not called by NGS and three ATRX mutations not being detected by Sanger sequencing. Importantly, in samples with low tumor cell content, NGS was able to identify mutant alleles that were not detectable by traditional methods. Information derived from NGS data identified potential targets for experimental therapy in 37/47 (79 %) glioblastomas, 9/10 (90 %) pilocytic astrocytomas, and 5/14 (36 %) medulloblastomas in the prospective target discovery cohort. In conclusion, we present the settings for high-throughput, adaptive next-generation sequencing in routine neuropathology diagnostics. Such an approach will likely become highly valuable in the near future for treatment decision making, as more therapeutic targets emerge and genetic information enters the classification of brain tumors. More... »

PAGES

903-910

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00401-015-1519-8

DOI

http://dx.doi.org/10.1007/s00401-015-1519-8

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1014983828

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26671409


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