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@ARTICLE{Sahm:130463,
author = {F. Sahm$^*$ and D. Schrimpf$^*$ and D. Jones$^*$ and J.
Meyer$^*$ and A. Kratz$^*$ and D. Reuss$^*$ and D.
Capper$^*$ and C. Koelsche$^*$ and A. Korshunov$^*$ and B.
P. O. Wiestler$^*$ and I. Buchhalter$^*$ and T. Milde$^*$
and F. Selt$^*$ and D. Sturm$^*$ and M. Kool$^*$ and M.
Hummel$^*$ and M. Bewerunge-Hudler$^*$ and C. Mawrin and U.
Schüller and C. Jungk and A. Wick and O. Witt$^*$ and M.
Platten$^*$ and C. Herold-Mende and A. Unterberg and S.
Pfister$^*$ and W. Wick$^*$ and A. von Deimling$^*$},
title = {{N}ext-generation sequencing in routine brain tumor
diagnostics enables an integrated diagnosis and identifies
actionable targets.},
journal = {Acta neuropathologica},
volume = {131},
number = {6},
issn = {1432-0533},
address = {Berlin},
publisher = {Springer},
reportid = {DKFZ-2017-05542},
pages = {903 - 910},
year = {2016},
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.},
cin = {G340 / G380 / B062 / G370 / G200 / C060 / W110 / G160 /
L101},
ddc = {610},
cid = {I:(DE-He78)G340-20160331 / I:(DE-He78)G380-20160331 /
I:(DE-He78)B062-20160331 / I:(DE-He78)G370-20160331 /
I:(DE-He78)G200-20160331 / I:(DE-He78)C060-20160331 /
I:(DE-He78)W110-20160331 / I:(DE-He78)G160-20160331 /
I:(DE-He78)L101-20160331},
pnm = {317 - Translational cancer research (POF3-317)},
pid = {G:(DE-HGF)POF3-317},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:26671409},
doi = {10.1007/s00401-015-1519-8},
url = {https://inrepo02.dkfz.de/record/130463},
}