DNA damage and nucleotide excision repair capacity in healthy individuals

Slyšková, Jana; Naccarati, Alessio; Polakova, Veronika; Pardini, Barbara; Vodičková, Ľudmila; Štětina, Rudolf; Schmuczerova, Jana; Šmerhovský, Zdeněk; Lipská, Ludmila; Vodička, Pavel

doi:10.1002/em.20650

Cited by 49 publications

(35 citation statements)

References 66 publications

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“…In the present study, XPD genotype did not influence the outcome of DNA damage assessed by SCGE or DNA repair as assessed by 8-oxo-dG, though it should be noted that 8-oxo-dG is not a specific product of the protein coded from XPD. These results are consistent with a recent study that reported that endogenous DNA damage and (+)-anti-benzo[a]-pyrene-7,8-dihydrodiol-9,10-epoxide-induced DNA repair capacity in PBMCs from a healthy population did not correlate with XPD751 genotypes [75]. …”

Section: Discussionsupporting

confidence: 93%

Assessment of DNA damage and repair in adults consuming allyl isothiocyanate or Brassica vegetables

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Clevidence

Albaugh

et al. 2013

The Journal of Nutritional Biochemistry

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Allylisothiocyanate (AITC) is a dietary component with possible anti-cancer effects, though much information about AITC and cancer has been obtained from cell studies. To investigate the effect of AITC on DNA integrity in vivo, a crossover study was conducted. Adults (n=46) consumed AITC, AITC-rich vegetables (mustard and cabbage), or a control treatment with a controlled diet for 10 days each. On day 11, volunteers provided blood and urine before and after consuming treatments. Volunteers were characterized for genotype for GSTM1 and GSTT1 (glutathione S-transferases) and XPD (DNA repair). DNA integrity in peripheral blood mononuclear cells (PBMCs) was assessed by single cell gel electrophoresis. Urine was analyzed for 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxodG) and creatinine. Ten day intake of neither AITC nor mustard/cabbage(M/C) resulted in statistically significant differences in DNA strand breaks (LS mean % DNA in tail ± SEM: 4.8 ± 0.6 for control, 5.7 ± 0.7 for AITC, 5.3 ± 0.6 for M/C) or urinary 8-oxodG (LS mean µg 8-oxodG/g creatinine ± SEM: 2.95 ± 0.09 for control, 2.88 ± 0.09 for AITC, 3.06 ± 0.09 for M/C). Both AITC and M/C increased DNA strand breaks 3h post-consumption (LS mean % DNA in tail ± SEM: 3.2 ± 0.7 for control, 8.3 ± 1.7 for AITC, 8.0 ± 1.7 for M/C), and this difference disappeared at 6h (4.2 ± 0.9 for control, 5.7 ± 1.2 for AITC, 5.5 ± 1.2 for M/C). Genotypes for GSTM1, GSTT1, and XPD were not associated with treatment effects. In summary, DNA damage appeared to be induced in the short term by AITC and AITC-rich products, but that damage disappeared quickly, and neither AITC nor AITC-rich products affected DNA base excision repair.

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Section: Discussionsupporting

confidence: 93%

Assessment of DNA damage and repair in adults consuming allyl isothiocyanate or Brassica vegetables

Charron

Clevidence

Albaugh

et al. 2013

The Journal of Nutritional Biochemistry

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“…We therefore also investigated whether DNA repair parameters assayed in this study are affected by gender. Gender effects on the level of endogenous SSBs have previously been reported in human PBMCs (Hofer et al , 2006; Slyskova et al , 2011), and indications of gender effects on DSBR were reported in a small study of 20 study subjects (Mayer et al , 1991). Several reports have suggested gender effects on other DNA repair pathways, primarily NER (Wei et al , 1993; Slyskova et al , 2011; Uppstad et al , 2011).…”

Section: Introductionmentioning

confidence: 77%

“…Higher levels of endogenous SSBs have been reported in PBMCs from men compared with women (Hofer et al , 2006; Slyskova et al , 2011). Men had higher levels of DNA damage than women in these studies including approximately 100 study subjects.…”

Section: Discussionmentioning

confidence: 97%

“…Gender differences have also been reported for the NER pathway investigating blood samples or human cell lines (Wei et al , 1993; Uppstad et al , 2011). But recently, a powerful epidemiological study investigating NER activity in PBMCs from 100 healthy individuals identified no statistically significant gender effect ( P = 0.86) (Slyskova et al , 2011). Similarly, none of the DNA repair parameters investigated in our study revealed significant gender differences (Table 2).…”

Section: Discussionmentioning

confidence: 99%

“…Some reports have suggested an age-related increase of endogenous SSBs in human PBMCs (Mutlu-Turkoglu et al , 2003; Moller, 2006; Humphreys et al , 2007), whereas others have suggested no age effect (Slyskova et al , 2011). An age-related decline of NHEJ functions in brain tissue has been suggested from animal studies (Ren & Pena de Ortiz, 2002; Vyjayanti & Rao, 2006).…”

Section: Introductionmentioning

confidence: 99%

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Age and gender effects on DNA strand break repair in peripheral blood mononuclear cells

et al. 2012

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Summary Exogenous and endogenous damage to DNA is constantly challenging the stability of our genome. This DNA damage increase the frequency of errors in DNA replication, thus causing point mutations or chromosomal rearrangements and has been implicated in aging, cancer, and neurodegenerative diseases. Therefore, efficient DNA repair is vital for the maintenance of genome stability. The general notion has been that DNA repair capacity decreases with age although there are conflicting results. Here, we focused on potential age-associated changes in DNA damage response and the capacities of repairing DNA single-strand breaks (SSBs) and double-strand breaks (DSBs) in human peripheral blood mononuclear cells (PBMCs). Of these lesions, DSBs are the least frequent but the most dangerous for cells. We have measured the level of endogenous SSBs, SSB repair capacity, γ-H2AX response, and DSB repair capacity in a study population consisting of 216 individuals from a population-based sample of twins aged 40– 77 years. Age in this range did not seem to have any effect on the SSB parameters. However, γ-H2AX response and DSB repair capacity decreased with increasing age, although the associations did not reach statistical significance after adjustment for batch effect across multiple experiments. No gender differences were observed for any of the parameters analyzed. Our findings suggest that in PBMCs, the repair of SSBs is maintained until old age, whereas the response to and the repair of DSBs decrease.

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High‐throughput measurement of double strand break global repair phenotype in peripheral blood mononuclear cells after long‐term cryopreservation

et al. 2023

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Peripheral blood mononuclear cells (PBMCs) are a useful model for biochemical assays, particularly for etiological studies. We describe here a method for measuring DNA repair capacity (DRC) in archival cryogenically preserved PBMCs. To model DRC, we measured γ-H2AX repair kinetics in thawed PBMCs after irradiation with 3 Gy gamma rays. Time-dependent fluorescently labeled γ-H2AX levels were measured at five time points from 1 to 20 h, yielding an estimate of global DRC repair kinetics as well as a measure of unrepaired double strand breaks at 20 h. While γ-H2AX levels are traditionally measured by either microscopy or flow-cytometry, we developed a protocol for imaging flow cytometry (IFC) that combines the detailed information of microscopy with the statistical power of flow methods. The visual imaging component of the IFC allows for monitoring aspects such as cellular health and apoptosis as well as fluorescence localization of the γ-H2AX signal, which ensures the power and significance of this technique. Application of a machinelearning based image classification improved flow cytometry fluorescent measurements by identifying apoptotic cells unable to undergo DNA repair. We present here DRC repair parameters from 18 frozen archival PBMCs and 28 fresh blood samples collected from a demographically diverse cohort of women measured in a highthroughput IFC format. This thaw method and assay can be used alone or in conjunction with other assays to measure etiological phenotypes in cryogenic biobanks of PBMCs.

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DNA damage and nucleotide excision repair capacity in healthy individuals

Cited by 49 publications

References 66 publications

Assessment of DNA damage and repair in adults consuming allyl isothiocyanate or Brassica vegetables

Assessment of DNA damage and repair in adults consuming allyl isothiocyanate or Brassica vegetables

Age and gender effects on DNA strand break repair in peripheral blood mononuclear cells

High‐throughput measurement of double strand break global repair phenotype in peripheral blood mononuclear cells after long‐term cryopreservation

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