Clonal expansion of mtDNA deletions: different disease models assessed by digital droplet PCR in single muscle cells.
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Authors
Trifunov, Selena
Pyle, Angela
Valentino, Maria Lucia
Liguori, Rocco
Yu-Wai-Man, Patrick
Burté, Florence
Duff, Jennifer
Diebold, Isabel
Rugolo, Michela
Horvath, Rita
Carelli, Valerio
Publication Date
2018-08-03Journal Title
Sci Rep
ISSN
2045-2322
Publisher
Springer Science and Business Media LLC
Volume
8
Issue
1
Pages
11682
Language
eng
Type
Article
Physical Medium
Electronic
Metadata
Show full item recordCitation
Trifunov, S., Pyle, A., Valentino, M. L., Liguori, R., Yu-Wai-Man, P., Burté, F., Duff, J., et al. (2018). Clonal expansion of mtDNA deletions: different disease models assessed by digital droplet PCR in single muscle cells.. Sci Rep, 8 (1), 11682. https://doi.org/10.1038/s41598-018-30143-z
Abstract
Deletions in mitochondrial DNA (mtDNA) are an important cause of human disease and their accumulation has been implicated in the ageing process. As mtDNA is a high copy number genome, the coexistence of deleted and wild-type mtDNA molecules within a single cell defines heteroplasmy. When deleted mtDNA molecules, driven by intracellular clonal expansion, reach a sufficiently high level, a biochemical defect emerges, contributing to the appearance and progression of clinical pathology. Consequently, it is relevant to determine the heteroplasmy levels within individual cells to understand the mechanism of clonal expansion. Heteroplasmy is reflected in a mosaic distribution of cytochrome c oxidase (COX)-deficient muscle fibers. We applied droplet digital PCR (ddPCR) to single muscle fibers collected by laser-capture microdissection (LCM) from muscle biopsies of patients with different paradigms of mitochondrial disease, characterized by the accumulation of single or multiple mtDNA deletions. By combining these two sensitive approaches, ddPCR and LCM, we document different models of clonal expansion in patients with single and multiple mtDNA deletions, implicating different mechanisms and time points for the development of COX deficiency in these molecularly distinct mitochondrial cytopathies.
Keywords
Adolescent, Adult, Aged, Biopsy, DNA, Mitochondrial, Electron Transport Complex IV, Female, GTP Phosphohydrolases, Gene Dosage, Genes, Recessive, Humans, Male, Middle Aged, Muscle Cells, Muscle Fibers, Skeletal, Mutation, Oxidative Phosphorylation, Polymerase Chain Reaction, Reproducibility of Results, Sequence Deletion, Succinate Dehydrogenase, Young Adult
Sponsorship
Wellcome Trust (109915_A_15_Z)
Medical Research Council (MR/N025431/2)
Identifiers
External DOI: https://doi.org/10.1038/s41598-018-30143-z
This record's URL: https://www.repository.cam.ac.uk/handle/1810/283111
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