Multisite de novo mutations in human offspring after paternal exposure to ionizing radiation

Holtgrewe, Manuel; Knaus, Alexej; Hildebrand, Gabriele; Pantel, Jean Tori; Santos, Miguel Rodríguez de los; Neveling, Kornelia; Goldmann, Jakob M.; Schubach, Max; Jäger, Marten; Coutelier, Marie; Mundlos, Stefan; Beule, Dieter; Sperling, Karl; Krawitz, Peter

doi:10.1038/s41598-018-33066-x

Cited by 25 publications

(15 citation statements)

References 26 publications

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“…Multisite mutations are believed to result from clustered lesions caused by IR 26,31 . A recent human study indicated that de novo multisite mutations were increased in the offspring of fathers who worked with radar units 32 .…”

Section: Discussionmentioning

confidence: 99%

Characteristics of induced mutations in offspring derived from irradiated mouse spermatogonia and mature oocytes

Satoh¹,

Asakawa²,

Nishimura³

et al. 2020

Sci Rep

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The exposure of germ cells to radiation introduces mutations in the genomes of offspring, and a previous whole-genome sequencing study indicated that the irradiation of mouse sperm induces insertions/deletions (indels) and multisite mutations (clustered single nucleotide variants and indels). However, the current knowledge on the mutation spectra is limited, and the effects of radiation exposure on germ cells at stages other than the sperm stage remain unknown. Here, we performed whole-genome sequencing experiments to investigate the exposure of spermatogonia and mature oocytes. We compared de novo mutations in a total of 24 F1 mice conceived before and after the irradiation of their parents. The results indicated that radiation exposure, 4 Gy of gamma rays, induced 9.6 indels and 2.5 multisite mutations in spermatogonia and 4.7 indels and 3.1 multisite mutations in mature oocytes in the autosomal regions of each F1 individual. Notably, we found two types of deletions, namely, small deletions (mainly 1~12 nucleotides) in non-repeat sequences, many of which showed microhomology at the breakpoint junction, and single-nucleotide deletions in mononucleotide repeat sequences. The results suggest that these deletions and multisite mutations could be a typical signature of mutations induced by parental irradiation in mammals.

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Section: Discussionmentioning

confidence: 99%

Characteristics of induced mutations in offspring derived from irradiated mouse spermatogonia and mature oocytes

Satoh¹,

Asakawa²,

Nishimura³

et al. 2020

Sci Rep

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“…Although there was a marginal increase in de novo mutations in the children conceived after therapy, this could be accounted for by the well-documented linear increase due to advancing paternal age (Kong et al 2012). Similarly, a WGS study of a non-Hodgkin lymphoma patient treated with radiotherapy and 12 soldiers working on equipment that was emitting high-frequency radiation found that the de novo point mutation load in their offspring was comparable to that of controls, although there appeared to be an increase in tandem and clustered point mutations (Holtgrewe et al 2018). Overall these reports, which suggest that cancer treatment has little long-term overall mutagenic effect on the male germline, should be reassuring, but it is important to recognise that the available studies remain small in scale, have a low resolution to quantify point mutations, and have only assessed the impact of a few treatment regimens.…”

Section: Introductionmentioning

confidence: 98%

The impact of chemo- and radiotherapy treatments on selfish de novo FGFR2 mutations in sperm of cancer survivors

et al. 2019

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STUDY QUESTION What effect does cancer treatment have on levels of spontaneous selfish fibroblast growth factor receptor 2 (FGFR2) point mutations in human sperm? SUMMARY ANSWER Chemotherapy and radiotherapy do not increase levels of spontaneous FGFR2 mutations in sperm but, unexpectedly, highly-sterilizing treatments dramatically reduce the levels of the disease-associated c.755C > G (Apert syndrome) mutation in sperm. WHAT IS KNOWN ALREADY Cancer treatments lead to short-term increases in gross DNA damage (chromosomal abnormalities and DNA fragmentation) but the long-term effects, particularly at the single nucleotide resolution level, are poorly understood. We have exploited an ultra-sensitive assay to directly quantify point mutation levels at the FGFR2 locus. STUDY DESIGN, SIZE, DURATION ‘Selfish’ mutations are disease-associated mutations that occur spontaneously in the sperm of most men and their levels typically increase with age. Levels of mutations at c.752–755 of FGFR2 (including c.755C > G and c.755C > T associated with Apert and Crouzon syndromes, respectively) in semen post-cancer treatment from 18 men were compared to levels in pre-treatment samples from the same individuals (n = 4) or levels in previously screened population controls (n = 99). PARTICIPANTS/MATERIALS, SETTING, METHODS Cancer patients were stratified into four different groups based on the treatments they received and the length of time for spermatogenesis recovery. DNA extracted from semen samples was analysed using a previously established highly sensitive assay to identify mutations at positions c.752–755 of FGFR2. Five to ten micrograms of semen genomic DNA was spiked with internal controls for quantification purposes, digested with MboI restriction enzyme and gel extracted. Following PCR amplification, further MboI digestion and a nested PCR with barcoding primers, samples were sequenced on Illumina MiSeq. Mutation levels were determined relative to the spiked internal control; in individuals heterozygous for a nearby common single nucleotide polymorphism (SNP), mutations were phased to their respective alleles. MAIN RESULTS AND THE ROLE OF CHANCE Patients treated with moderately-sterilizing alkylating regimens and who recovered spermatogenesis within <3 years after therapy (Group 3, n = 4) or non − alkylating chemotherapy and/or low gonadal radiation doses (Group 1, n = 4) had mutation levels similar to untreated controls. However, patients who had highly-sterilizing alkylating treatments (i.e. >5 years to spermatogenesis recovery) (Group 2, n = 7) or pelvic radiotherapy (Group 4, n = 3) exhibited c.755C > G mutation levels at or below background. Two patients (A and B) treated with highly-sterilizing alkylating agents demonstrated a clear reduction from pre-treatment levels; however pre-treatment samples were not available for the other patients with low mutation levels. Therefore, although based on their age we would expect detectable levels of mutations, we cannot exclude the possibility that these patients also had low mutation levels pre-treatment. In three patients with low c.755C > G levels at the first timepoint post-treatment, we observed increasing mutation levels over time. For two such patients we could phase the mutation to a nearby polymorphism (SNP) and determine that the mutation counts likely originated from a single or a small number of mutational events. LIMITATIONS, REASONS FOR CAUTION This study was limited to 18 patients with different treatment regimens; for nine of the 18 patients, samples from only one timepoint were available. Only 12 different de novo substitutions at the FGFR2 c.752–755 locus were assessed, two of which are known to be disease associated. WIDER IMPLICATIONS OF THE FINDINGS Our data add to the body of evidence from epidemiological studies and experimental data in humans suggesting that male germline stem cells are resilient to the accumulation of spontaneous mutations. Collectively, these data should provide physicians and health-care professionals with reassuring experimental-based evidence for counselling of male cancer patients contemplating their reproductive options several years after treatment. STUDY FUNDING/COMPETING INTEREST(S) This work was primarily supported by grants from the Wellcome (grant 091182 to AG and AOMW; grant 102 731 to AOMW), the University of Oxford Medical Sciences Division Internal Fund (grant 0005128 to GJM and AG), the National Institute for Health Research (NIHR) Oxford Biomedical Research Centre Programme (to AG) and the US National Institutes of Health (to MLM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. None of the authors has any conflicts of interest to declare. TRIAL REGISTRATION NUMBER NA

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“…Some CM are the result of so-called de novo mutations which arise in the population. De novo mutations arise during gametogenesis or during early embryonic development and can determine the expression of CM such as polydactyly, LRD and MCM in offspring ( Lazyuk et al., 1999 ; Yablokov et al., 2016 ; Holtgrewe et al., 2018 ). It was found that these de novo CM are found significantly more often in areas with a concentration of Cs-137 contamination of more than 555 kBq/m 2 ( Lazyuk et al., 1999 ; Yablokov et al., 2016 ).…”

Section: Introductionmentioning

confidence: 99%

De novo congenital malformation frequencies in children from the Bryansk region following the Chernobyl disaster (2000–2017)

Geger

Lagerev

Pugach

et al. 2020

Heliyon

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Background Ionizing radiation and chemical pollution can disrupt normal embryonic development and lead to congenital malformations and fetal death. We used official government statistical data for 2000–2017 to test the hypothesis that radioactive and chemical pollutants influenced the frequency of de novo congenital malformations in newborns of the Bryansk region of southwest Russia. Methods A variety of statistical approaches were used to assess congenital malformation frequencies including the Shapiro-Wilk test, White's homoscedasticity test, Wilcoxon T-test, Spearman's rank correlation test, and the inversely proportional regression. Results We found that the frequency of polydactyly, multiple congenital malformations, and the frequency of de novo congenital malformations in newborns were significantly higher (p = 0.001–0.054) in regions with elevated radioactive, chemical and combined contamination. Polydactyly, multiple congenital malformations, and the sum of all congenital malformations were 4.7–7.4 times, 2.5–6.8 times, and 3.5–4.6 times higher in contaminated regions in comparison with the control group. The combination of both radioactive and chemical pollutants led to significantly higher frequencies of multiple congenital malformations when compared to regions with only one pollutant (radiation alone: 2.2 times, p = 0.034; chemical pollutants alone: 1.9 times, p = 0.008) implying that the effects of these stressors were at minimum additive. Although there was a trend for decreasing frequencies of multiple congenital malformations during the 2000–2017 period in areas of combined pollution, the opposite was true for regions with radioactive or chemical pollutants alone. However, overall, our models suggest that the frequency of multiple congenital malformations in areas of combined pollution will significantly (p = 0.027) exceed the frequencies observed for regions containing radioactive or chemical pollutants alone by 39.6% and 45.7% respectively, by 2018–2023. Conclusion These findings suggest additive and potentially synergistic effects of radioactive and chemical pollutants on the frequencies of multiple congenital malformations in the Bryansk region of southwestern Russia.

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Multisite de novo mutations in human offspring after paternal exposure to ionizing radiation

Cited by 25 publications

References 26 publications

Characteristics of induced mutations in offspring derived from irradiated mouse spermatogonia and mature oocytes

Characteristics of induced mutations in offspring derived from irradiated mouse spermatogonia and mature oocytes

The impact of chemo- and radiotherapy treatments on selfish de novo FGFR2 mutations in sperm of cancer survivors

De novo congenital malformation frequencies in children from the Bryansk region following the Chernobyl disaster (2000–2017)

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