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@ARTICLE{Broche:304252,
author = {J. Broche$^*$ and O. Kelemen and A. Sekar and L. Schütz
and F. Muyas and A. Forschner and C. Schroeder$^*$ and S.
Ossowski$^*$},
title = {{G}ene{B}its: ultra-sensitive tumour-informed ct{DNA}
monitoring of treatment response and relapse in cancer
patients.},
journal = {Journal of translational medicine},
volume = {23},
number = {1},
issn = {1479-5876},
address = {London},
publisher = {BioMed Central},
reportid = {DKFZ-2025-01806},
pages = {964},
year = {2025},
abstract = {Circulating tumour DNA (ctDNA) in liquid biopsies has
emerged as a powerful biomarker in cancer patients. Its
relative abundance in cell-free DNA serves as a proxy for
the overall tumour burden. Here we present GeneBits, a
method for cancer therapy monitoring and relapse detection.
GeneBits employs tumour-informed enrichment panels targeting
20-100 somatic single-nucleotide variants (SNVs) in
plasma-derived DNA, combined with ultra-deep sequencing and
unique molecular barcoding. In conjunction with the newly
developed computational method umiVar, GeneBits enables
accurate detection of molecular residual disease and early
relapse identification.To assess the performance of GeneBits
and umiVar, we conducted benchmarking experiments using
three different commercial cell-free DNA reference
standards. These standards were tested with targeted
next-generation sequencing (NGS) workflows from both IDT and
Twist, allowing us to evaluate the consistency and accuracy
of our approach across different oligo-enrichment
strategies. GeneBits achieved comparable depth of coverage
across all target sites, demonstrating robust performance
independent of the enrichment kit used. For duplex reads
with ≥ 4x UMI-family size, umiVar achieved exceptionally
low error rates, ranging from 7.4×10-7 to 7.5×10-5. Even
when including mixed consensus reads (duplex $\&$ simplex),
error rates remained low, between 6.1×10-6 and 9×10-5.
Furthermore, umiVar enabled variant detection at a limit of
detection as low as $0.0017\%,$ with no false positive calls
in mutation-free reference samples. In a reanalysed melanoma
cohort, variant allele frequency kinetics closely mirrored
imaging results, confirming the clinical relevance of our
method.GeneBits and umiVar enable highly accurate therapy
and relapse monitoring in plasma as well as identification
of molecular residual disease within four weeks of tumour
surgery or biopsy. By leveraging small, tumour-informed
sequencing panels, GeneBits provides a targeted,
cost-effective, and scalable approach for ctDNA-based cancer
monitoring. The benchmarking experiments using multiple
commercial cell-free DNA reference standards confirmed the
high sensitivity and specificity of GeneBits and umiVar,
making them valuable tools for precision oncology. UmiVar is
available at https://github.com/imgag/umiVar .},
keywords = {Humans / Circulating Tumor DNA: genetics / Circulating
Tumor DNA: blood / Neoplasms: genetics / Neoplasms: therapy
/ Neoplasms: blood / High-Throughput Nucleotide Sequencing /
Neoplasm Recurrence, Local: genetics / Treatment Outcome /
Recurrence / Liquid Biopsy / Polymorphism, Single
Nucleotide: genetics / Cell-free DNA (Other) / Circulating
tumour DNA (Other) / Liquid biopsy (Other) / Molecular
residual disease (Other) / Next-Generation-Sequencing
(Other) / Relapse detection (Other) / Treatment monitoring
(Other) / Ultra-deep sequencing (Other) / Unique molecular
barcode (Other) / Circulating Tumor DNA (NLM Chemicals)},
cin = {TU01},
ddc = {610},
cid = {I:(DE-He78)TU01-20160331},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40866952},
doi = {10.1186/s12967-025-06993-3},
url = {https://inrepo02.dkfz.de/record/304252},
}