CometChip analysis of human primary lymphocytes enables quantification of inter-individual differences in the kinetics of repair of oxidative DNA damage

Free Radic Biol Med. 2021 Oct;174:89-99. doi: 10.1016/j.freeradbiomed.2021.07.033. Epub 2021 Jul 26.

Abstract

Although DNA repair is known to impact susceptibility to cancer and other diseases, relatively few population studies have been performed to evaluate DNA repair kinetics in people due to the difficulty of assessing DNA repair in a high-throughput manner. Here we use the CometChip, a high-throughput comet assay, to explore inter-individual variation in repair of oxidative damage to DNA, a known risk factor for aging, cancer and other diseases. DNA repair capacity after H2O2-induced DNA oxidation damage was quantified in peripheral blood mononuclear cells (PBMCs). For 10 individuals, blood was drawn at several times over the course of 4-6 weeks. In addition, blood was drawn once from each of 56 individuals. DNA damage levels were quantified prior to exposure to H2O2 and at 0, 15, 30, 60, and 120-min post exposure. We found that there is significant variability in DNA repair efficiency among individuals. When subdivided into quartiles by DNA repair efficiency, we found that the average t1/2 is 81 min for the slowest group and 24 min for the fastest group. This work shows that the CometChip can be used to uncover significant differences in repair kinetics among people, pointing to its utility in future epidemiological and clinical studies.

Keywords: CometChip; DNA damage; DNA repair; Repair kinetics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Comet Assay
  • DNA Damage
  • DNA Repair
  • Humans
  • Hydrogen Peroxide*
  • Individuality
  • Kinetics
  • Leukocytes, Mononuclear*
  • Lymphocytes
  • Oxidative Stress / genetics

Substances

  • Hydrogen Peroxide