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Somatic mutation landscapes at single-molecule resolution.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Harvey, Luke MR 
Mitchell, Emily 
Lensing, Stefanie V 

Abstract

Somatic mutations drive the development of cancer and may contribute to ageing and other diseases1,2. Despite their importance, the difficulty of detecting mutations that are only present in single cells or small clones has limited our knowledge of somatic mutagenesis to a minority of tissues. Here, to overcome these limitations, we developed nanorate sequencing (NanoSeq), a duplex sequencing protocol with error rates of less than five errors per billion base pairs in single DNA molecules from cell populations. This rate is two orders of magnitude lower than typical somatic mutation loads, enabling the study of somatic mutations in any tissue independently of clonality. We used this single-molecule sensitivity to study somatic mutations in non-dividing cells across several tissues, comparing stem cells to differentiated cells and studying mutagenesis in the absence of cell division. Differentiated cells in blood and colon displayed remarkably similar mutation loads and signatures to their corresponding stem cells, despite mature blood cells having undergone considerably more divisions. We then characterized the mutational landscape of post-mitotic neurons and polyclonal smooth muscle, confirming that neurons accumulate somatic mutations at a constant rate throughout life without cell division, with similar rates to mitotically active tissues. Together, our results suggest that mutational processes that are independent of cell division are important contributors to somatic mutagenesis. We anticipate that the ability to reliably detect mutations in single DNA molecules could transform our understanding of somatic mutagenesis and enable non-invasive studies on large-scale cohorts.

Description

Keywords

Alzheimer Disease, Blood Cells, Cell Differentiation, Cell Division, Cohort Studies, Colon, DNA Mutational Analysis, Epithelium, Granulocytes, Healthy Volunteers, Humans, Male, Middle Aged, Muscle, Smooth, Mutagenesis, Mutation, Mutation Rate, Neurons, Single Molecule Imaging, Stem Cells

Journal Title

Nature

Conference Name

Journal ISSN

0028-0836
1476-4687

Volume Title

593

Publisher

Springer Science and Business Media LLC

Rights

All rights reserved
Sponsorship
Wellcome Trust (107630/Z/15/Z)
Wellcome Trust (203151/Z/16/Z)
Bloodwise (15008)
European Research Council (715371)
Medical Research Council (MC_PC_17230)