Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses

Abstract De novo mutations (DNMs) are drivers of genetic disorders. However, the study of DNMs is hampered by technological limitations preventing accurate quantification of ultra-rare mutations. Duplex Sequencing (DS) theoretically has < 1 error/billion base-pairs (bp). To determine the DS utili...

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Main Authors: Jonatan Axelsson, Danielle LeBlanc, Habiballah Shojaeisaadi, Matthew J Meier, Devon M. Fitzgerald, Daniela Nachmanson, Jedidiah Carlson, Alexandra Golubeva, Jake Higgins, Thomas Smith, Fang Yin Lo, Richard Pilsner, Andrew Williams, Jesse Salk, Francesco Marchetti, Carole Yauk
Format: Article
Language:English
Published: Nature Portfolio 2024-10-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-024-73587-2
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author Jonatan Axelsson
Danielle LeBlanc
Habiballah Shojaeisaadi
Matthew J Meier
Devon M. Fitzgerald
Daniela Nachmanson
Jedidiah Carlson
Alexandra Golubeva
Jake Higgins
Thomas Smith
Fang Yin Lo
Richard Pilsner
Andrew Williams
Jesse Salk
Francesco Marchetti
Carole Yauk
author_facet Jonatan Axelsson
Danielle LeBlanc
Habiballah Shojaeisaadi
Matthew J Meier
Devon M. Fitzgerald
Daniela Nachmanson
Jedidiah Carlson
Alexandra Golubeva
Jake Higgins
Thomas Smith
Fang Yin Lo
Richard Pilsner
Andrew Williams
Jesse Salk
Francesco Marchetti
Carole Yauk
author_sort Jonatan Axelsson
collection DOAJ
description Abstract De novo mutations (DNMs) are drivers of genetic disorders. However, the study of DNMs is hampered by technological limitations preventing accurate quantification of ultra-rare mutations. Duplex Sequencing (DS) theoretically has < 1 error/billion base-pairs (bp). To determine the DS utility to quantify and characterize DNMs, we analyzed DNA from blood and spermatozoa from six healthy, 18-year-old Swedish men using the TwinStrand DS mutagenesis panel (48 kb spanning 20 genic and intergenic loci). The mean single nucleotide variant mutation frequency (MF) was 1.2 × 10− 7 per bp in blood and 2.5 × 10− 8 per bp in sperm, with the most common base substitution being C > T. Blood MF and substitution spectrum were similar to those reported in blood cells with an orthogonal method. The sperm MF was in the same order of magnitude and had a strikingly similar spectrum to DNMs from publicly available whole genome sequencing data from human pedigrees (1.2 × 10− 8 per bp). DS revealed much larger numbers of insertions and deletions in sperm over blood, driven by an abundance of putative extra-chromosomal circular DNAs. The study indicates the strong potential of DS to characterize human DNMs to inform factors that contribute to disease susceptibility and heritable genetic risks.
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spelling doaj-art-642eb87b6f244a7fb80c1b98bc0a85492024-12-22T12:26:03ZengNature PortfolioScientific Reports2045-23222024-10-0114111510.1038/s41598-024-73587-2Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analysesJonatan Axelsson0Danielle LeBlanc1Habiballah Shojaeisaadi2Matthew J Meier3Devon M. Fitzgerald4Daniela Nachmanson5Jedidiah Carlson6Alexandra Golubeva7Jake Higgins8Thomas Smith9Fang Yin Lo10Richard Pilsner11Andrew Williams12Jesse Salk13Francesco Marchetti14Carole Yauk15Department of Biology, University of OttawaEnvironmental Health Science and Research Bureau, Health CanadaEnvironmental Health Science and Research Bureau, Health CanadaEnvironmental Health Science and Research Bureau, Health CanadaTwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.TwinStrand Biosciences, Inc.Department of Obstetrics & Gynecology, Wayne State UniversityEnvironmental Health Science and Research Bureau, Health CanadaTwinStrand Biosciences, Inc.Environmental Health Science and Research Bureau, Health CanadaDepartment of Biology, University of OttawaAbstract De novo mutations (DNMs) are drivers of genetic disorders. However, the study of DNMs is hampered by technological limitations preventing accurate quantification of ultra-rare mutations. Duplex Sequencing (DS) theoretically has < 1 error/billion base-pairs (bp). To determine the DS utility to quantify and characterize DNMs, we analyzed DNA from blood and spermatozoa from six healthy, 18-year-old Swedish men using the TwinStrand DS mutagenesis panel (48 kb spanning 20 genic and intergenic loci). The mean single nucleotide variant mutation frequency (MF) was 1.2 × 10− 7 per bp in blood and 2.5 × 10− 8 per bp in sperm, with the most common base substitution being C > T. Blood MF and substitution spectrum were similar to those reported in blood cells with an orthogonal method. The sperm MF was in the same order of magnitude and had a strikingly similar spectrum to DNMs from publicly available whole genome sequencing data from human pedigrees (1.2 × 10− 8 per bp). DS revealed much larger numbers of insertions and deletions in sperm over blood, driven by an abundance of putative extra-chromosomal circular DNAs. The study indicates the strong potential of DS to characterize human DNMs to inform factors that contribute to disease susceptibility and heritable genetic risks.https://doi.org/10.1038/s41598-024-73587-2Sperm DNA mutationsDe novo mutationsMutation frequencyMutational spectrumExtrachromosomal circular DNADuplex sequencing
spellingShingle Jonatan Axelsson
Danielle LeBlanc
Habiballah Shojaeisaadi
Matthew J Meier
Devon M. Fitzgerald
Daniela Nachmanson
Jedidiah Carlson
Alexandra Golubeva
Jake Higgins
Thomas Smith
Fang Yin Lo
Richard Pilsner
Andrew Williams
Jesse Salk
Francesco Marchetti
Carole Yauk
Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
Scientific Reports
Sperm DNA mutations
De novo mutations
Mutation frequency
Mutational spectrum
Extrachromosomal circular DNA
Duplex sequencing
title Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
title_full Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
title_fullStr Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
title_full_unstemmed Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
title_short Frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio-based de novo mutation analyses
title_sort frequency and spectrum of mutations in human sperm measured using duplex sequencing correlate with trio based de novo mutation analyses
topic Sperm DNA mutations
De novo mutations
Mutation frequency
Mutational spectrum
Extrachromosomal circular DNA
Duplex sequencing
url https://doi.org/10.1038/s41598-024-73587-2
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