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Mitochondrial DNA deletion mutations increase exponentially with age in human skeletal muscle.
Herbst, Allen; Lee, Cathy C; Vandiver, Amy R; Aiken, Judd M; McKenzie, Debbie; Hoang, Austin; Allison, David; Liu, Nianjun; Wanagat, Jonathan.
Afiliación
  • Herbst A; Department of Agricultural, Food and Nutritional Sciences, University of Alberta, Edmonton, AB, Canada.
  • Lee CC; Veterans Administration Greater Los Angeles Healthcare System, Los Angeles, USA.
  • Vandiver AR; Division of Geriatrics, Department of Medicine, UCLA, Los Angeles, CA, USA.
  • Aiken JM; Division of Dermatology, Department of Medicine, UCLA, 10945 Le Conte Ave, Suite 2339, Los Angeles, CA, 90095, USA.
  • McKenzie D; Department of Agricultural, Food and Nutritional Sciences, University of Alberta, Edmonton, AB, Canada.
  • Hoang A; Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada.
  • Allison D; Division of Geriatrics, Department of Medicine, UCLA, Los Angeles, CA, USA.
  • Liu N; Department of Epidemiology and Biostatistics, Indiana University Bloomington, Bloomington, IN, USA.
  • Wanagat J; Department of Epidemiology and Biostatistics, Indiana University Bloomington, Bloomington, IN, USA.
Aging Clin Exp Res ; 33(7): 1811-1820, 2021 Jul.
Article en En | MEDLINE | ID: mdl-32965609
BACKGROUND: Mitochondrial DNA (mtDNA) deletion mutations lead to electron transport chain-deficient cells and age-induced cell loss in multiple tissues and mammalian species. Accurate quantitation of somatic mtDNA deletion mutations could serve as an index of age-induced cell loss. Quantitation of mtDNA deletion molecules is confounded by their low abundance in tissue homogenates, the diversity of deletion breakpoints, stochastic accumulation in single cells, and mosaic distribution between cells. AIMS: Translate a pre-clinical assay to quantitate mtDNA deletions for use in human DNA samples, with technical and biological validation, and test this assay on human subjects of different ages. METHODS: We developed and validated a high-throughput droplet digital PCR assay that quantitates human mtDNA deletion frequency. RESULTS: Analysis of human quadriceps muscle samples from 14 male subjects demonstrated that mtDNA deletion frequency increases exponentially with age-on average, a 98-fold increase from age 20-80. Sequence analysis of amplification products confirmed the specificity of the assay for human mtDNA deletion breakpoints. Titration of synthetic mutation mixtures found a lower limit of detection of at least 0.6 parts per million. Using muscle DNA from 6-month-old mtDNA mutator mice, we measured a 6.4-fold increase in mtDNA deletion frequency (i.e., compared to wild-type mice), biologically validating the approach. DISCUSSION/CONCLUSIONS: The exponential increase in mtDNA deletion frequency is concomitant with the known muscle fiber loss and accelerating mortality that occurs with age. The improved assay permits the accurate and sensitive quantification of deletion mutations from DNA samples and is sufficient to measure changes in mtDNA deletion mutation frequency in healthy individuals across the lifespan and, therefore, patients with suspected mitochondrial diseases.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN Mitocondrial / Músculo Esquelético Límite: Adult / Aged / Aged80 / Animals / Humans / Male / Middle aged Idioma: En Revista: Aging Clin Exp Res Asunto de la revista: GERIATRIA Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Alemania
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN Mitocondrial / Músculo Esquelético Límite: Adult / Aged / Aged80 / Animals / Humans / Male / Middle aged Idioma: En Revista: Aging Clin Exp Res Asunto de la revista: GERIATRIA Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Alemania