Error-corrected flow-based sequencing at whole-genome scale and its application to circulating cell-free DNA profiling.

TitleError-corrected flow-based sequencing at whole-genome scale and its application to circulating cell-free DNA profiling.
Publication TypeJournal Article
Year of Publication2025
AuthorsCheng APellan, Widman AJ, Arora A, Rusinek I, Sossin A, Rajagopalan S, Midler N, Hooper WF, Murray RM, Halmos D, Langanay T, Chu H, Inghirami G, Potenski C, Germer S, Marton M, Manaa D, Helland A, Furatero R, McClintock J, Winterkorn L, Steinsnyder Z, Wang Y, Alimohamed AI, Malbari MS, Saxena A, Callahan MK, Frederick DT, Spain L, Sigouros M, Manohar J, King A, Wilkes D, Otilano J, Elemento O, Mosquera JMiguel, Jaimovich A, Lipson D, Turajlic S, Zody MC, Altorki NK, Wolchok JD, Postow MA, Robine N, Faltas BM, Boland G, Landau DA
JournalNat Methods
Volume22
Issue5
Pagination973-981
Date Published2025 May
ISSN1548-7105
KeywordsCell-Free Nucleic Acids, Circulating Tumor DNA, Genomics, High-Throughput Nucleotide Sequencing, Humans, Melanoma, Mutation, Sequence Analysis, DNA, Whole Genome Sequencing
Abstract

Differentiating sequencing errors from true variants is a central genomics challenge, calling for error suppression strategies that balance costs and sensitivity. For example, circulating cell-free DNA (ccfDNA) sequencing for cancer monitoring is limited by sparsity of circulating tumor DNA, abundance of genomic material in samples and preanalytical error rates. Whole-genome sequencing (WGS) can overcome the low abundance of ccfDNA by integrating signals across the mutation landscape, but higher costs limit its wide adoption. Here, we applied deep (~120×) lower-cost WGS (Ultima Genomics) for tumor-informed circulating tumor DNA detection within the part-per-million range. We further leveraged lower-cost sequencing by developing duplex error-corrected WGS of ccfDNA, achieving 7.7 × 10 error rates, allowing us to assess disease burden in individuals with melanoma and urothelial cancer without matched tumor sequencing. This error-corrected WGS approach will have broad applicability across genomics, allowing for accurate calling of low-abundance variants at efficient cost and enabling deeper mapping of somatic mosaicism as an emerging central aspect of aging and disease.

DOI10.1038/s41592-025-02648-9
Alternate JournalNat Methods
PubMed ID40217113
PubMed Central IDPMC12077166
Grant ListR01 CA266619 / CA / NCI NIH HHS / United States
P30 CA08748 / / U.S. Department of Health & Human Services | NIH | National Cancer Institute (NCI) /
K08 CA263301 / CA / NCI NIH HHS / United States
FC10988 / / Wellcome Trust (Wellcome) /
P50 CA221745 / CA / NCI NIH HHS / United States
A29911, FC10988 / / Cancer Research UK (CRUK) /
R01-CA266619-01 / / U.S. Department of Health & Human Services | NIH | National Cancer Institute (NCI) /
/ WT_ / Wellcome Trust / United Kingdom
P30 CA008748 / CA / NCI NIH HHS / United States
U01 CA247439 / CA / NCI NIH HHS / United States