% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@ARTICLE{Taylor:306869,
author = {A. M. Taylor and J. T. Lombardi and A. J. Patel$^*$ and A.
Tamariz and J. Martin and M. J. Bookland and D. S. Hersh and
E. Cantor and X. Song and F. Sahm$^*$ and P. K. Ng and J. J.
Gell and C. C. Lau},
title = {{A} feasibility study of enzymatic methylation sequencing
of cell-free {DNA} from cerebrospinal fluid of pediatric
central nervous system tumor patients for molecular
classification.},
journal = {Neuro-oncology advances},
volume = {7},
number = {1},
issn = {2632-2498},
address = {Oxford},
publisher = {Oxford University Press},
reportid = {DKFZ-2025-02846},
pages = {vdaf159},
year = {2025},
abstract = {Array-based DNA methylation profiling is the gold standard
for central nervous system (CNS) tumor molecular
classification, but requires over 100 ng input DNA from
surgical tissue. Cell-free tumor DNA (cfDNA) in
cerebrospinal fluid (CSF) offers an alternative for
diagnosis and disease monitoring. This study aimed to test
the utilization of enzymatic DNA methylation sequencing
(EM-seq) methods to overcome input DNA limitations.We used
the NEBNext EM-seq v2 kit on various amounts of cfDNA, as
low as 0.1 ng, extracted from archival CSF samples of 10
patients with CNS tumors. Tumor classification was performed
via MNP-Flex using CpG sites overlapping those on the
MethylationEPIC array.EM-seq provided sufficient genomic
coverage for 10 and 1 ng input DNA samples to generate
global DNA methylation profiles. Samples with 0.1 ng input
showed lower coverage due to read duplication. Methylation
levels for CpG sites with at least 5× coverage were highly
correlated across various input DNA amounts, indicating that
lower input cfDNA can still be used for tumor
classification. The MNP-Flex classifier, trained on tissue
DNA methylation data, successfully predicted CNS tumor types
for 7 out of 10 CSF samples using EM-seq methylation data
with only 1 ng of input cfDNA, consistent with diagnoses
based on tissue MethylationEPIC classification and/or
histopathology. Additionally, we detected focal and
arm-level copy number alterations previously identified via
clinical cytogenetics of tumor tissue.This study
demonstrated the feasibility of CNS tumor molecular
classification based on CSF using the EM-seq approach, and
establishes potential sample quality limitations for future
studies.},
keywords = {CNS tumor classification (Other) / MNP-flex (Other) /
cell-free DNA (Other) / enzymatic methylation sequencing
(Other) / molecular diagnosis (Other)},
cin = {B062 / B300},
ddc = {610},
cid = {I:(DE-He78)B062-20160331 / I:(DE-He78)B300-20160331},
pnm = {312 - Funktionelle und strukturelle Genomforschung
(POF4-312)},
pid = {G:(DE-HGF)POF4-312},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40746948},
pmc = {pmc:PMC12311925},
doi = {10.1093/noajnl/vdaf159},
url = {https://inrepo02.dkfz.de/record/306869},
}