Inhibition of histone deacetylases enhances DNA damage repair in SCNT embryos

Bohrer, Rodrigo Camponogara; Duggavathi, Raj; Bordignon, Vilceu

doi:10.4161/cc.29215

Cited by 30 publications

(27 citation statements)

References 68 publications

(85 reference statements)

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“…Therefore, we hypothesized that CDK2 knockdown-induced cell cycle arrest may be caused by increased accumulation of DNA damage during early embryonic development. Indeed, by analyzing the number of γH2AX fluorescent foci [37,38], we demonstrated that CDK2 knockdown embryos have more DNA DSBs than control embryos on days 3 and 5 of development. In response to DNA damage, cell cycle checkpoints (G1, S, G2/M) are activated, stopping cell cycle progression to allow time for repair, thereby preventing transmission of damaged or incompletely replicated chromosomes [3].…”

Section: Discussionmentioning

confidence: 98%

CDK2 Is Required for the DNA Damage Response During Porcine Early Embryonic Development

Wang

Kim

2016

Biology of Reproduction

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Cyclin-dependent kinase (CDK) 2 inhibition plays a central role in DNA damage-induced cell cycle arrest and DNA repair. However, whether CDK2 also influences early porcine embryo development is unknown. In this study, we examined whether CDK2 is involved in the regulation of oocyte meiosis and early embryonic development of porcine embryos. We found that disrupting CDK2 activity with RNAi or an inhibitor did not affect meiotic resumption or meiosis II arrest. However, CDK2 inhibitor-treated embryos showed delayed cleavage and ceased development before the blastocyst stage. Disrupting CDK2 activity is able to induce sustained DNA damage, as demonstrated by the formation of distinct gammaH2AX foci in nuclei of Day-3 and Day-5 embryos. Inhibiting CDK2 triggers a DNA damage checkpoint by activation of the ataxia telangiectasia mutated (ATM)-P53-P21 pathway. However, the mRNA expression of genes involved in nonhomologous end joining or homologous recombination pathways for double-strand break repair were reduced after administering CDK2 inhibitor to 5-day-old embryos. Furthermore, CDK2 inhibition caused apoptosis in Day-7 blastocysts. Thus, our results indicate that an ATM-P53-P21 DNA damage checkpoint is intact in the absence of CDK2; however, CDK2 is important for proper repair of the damaged DNA by either directly or indirectly influencing DNA repair-related gene expression.

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Section: Discussionmentioning

confidence: 98%

CDK2 Is Required for the DNA Damage Response During Porcine Early Embryonic Development

Wang

Kim

2016

Biology of Reproduction

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“…Conversely, the HR pathway seems to be more important than the NHEJ pathway for DSB repair in embryonic stem cells (40). In early developing embryos, the two repair pathways are active (2, 41) and gene transcripts of the two pathways are up‐regulated in response to UV‐induced DNA damage (4, 6). Nonetheless, previous studies have not determined if both pathways play crucial roles in DSB repair during early embryo development, and whether there is a synergistic or compensatory activation of the response between the pathways.…”

Section: Discussionmentioning

confidence: 99%

“…The same software was used to determine the number of DSBs by counting the number of H2AX139ph foci >0.3 μm 3 in each nucleus (37). We also counted the number of fluorescent foci for 53BP1 and RAD51 that colocalized with H2AX139ph foci (6).…”

Section: Scntmentioning

confidence: 99%

Double‐strand DNA breaks are mainly repaired by the homologous recombination pathway in early developing swine embryos

et al. 2018

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DNA double-strand breaks (DSBs) are less frequent than single-strand breaks but have more harmful consequences on cell survival and physiology. Homologous recombination (HR) and nonhomologous end-joining (NHEJ) are the two main pathways that are responsible for DSB repair in eukaryotic cells, but their importance for the preservation of genome stability in totipotent blastomeres of early developing embryos has not been determined. In this study, we observed that the chemical inhibition of HR or both pathways, but not NHEJ alone, increased the number of DSBs, reduced embryo development to the blastocyst stage, and resulted in embryos with higher proportions of apoptotic cells. Targeted knockdown of ATM (ataxia telangiectasia mutated) and ATR (ataxia telangiectasia and Rad3 related; HR regulators) and DNA-dependent protein kinase (NHEJ regulator) mRNAs revealed that the attenuation of HR or both HR and NHEJ regulators severely impaired blastocyst formation and quality. Attenuation of ATM alone resulted in a higher incidence of DSBs, lower development and embryo quality, and increased mRNA abundance of genes that are involved in either repair pathway. These findings indicate that HR is the main pathway responsible for the promotion of DSB repair in early developing embryos, and that ATM seems to be more important than ATR in the regulation of the HR pathway in mammalian embryos.-Bohrer, R. C., Dicks, N., Gutierrez, K., Duggavathi, R., Bordignon, V. Double-strand DNA breaks are mainly repaired by the homologous recombination pathway in early developing swine embryos.

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“…Many studies have showed that the developmental defects of cloned embryos fail to eliminate epigeneticspecific markers completely and establish the related gamete markers themselves (Kishigami et al, 2006;Latham, 2005;Rideout et al, 2001;Shi et al, 2003). Moreover, many studies also found that an abnormal epigenetic modification might result in the developmental failure of SCNT embryos, such as abnormal histone acetylation and DNA methylation in reprogramming (Bohrer et al, 2014;Enright et al, 2005;Jafarpour et al, 2011;Sangalli et al, 2014). Histone acetylation is a part of epigenetic modification (Grunstein, 1997;Turner, 2002) and is related to transcriptional activation by regulating the chromosome structure.…”

Section: Discussionmentioning

confidence: 96%

Effects of Scriptaid on the Histone Acetylation, DNA Methylation and Development of Buffalo Somatic Cell Nuclear Transfer Embryos

Sun

Zhu

et al. 2015

Cellular Reprogramming

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The present study was undertaken to examine the effect of Scriptaid treatment on histone acetylation, DNA methylation, expression of genes related to histone acetylation, and development of buffalo somatic cell nuclear transfer (SCNT) embryos. Treatment of buffalo SCNT embryos with 500 nM Scriptaid for 24 h resulted in a significant increase in the blastocyst formation rate (28.2% vs. 13.6%, p<0.05). Meanwhile, treatment of buffalo SCNT embryos with Scriptaid also resulted in higher acetylation levels of H3K18 and lower methylation levels of global DNA at the blastocyst stage, which was similar to fertilized counterparts. The expression levels of CBP, p300, HAT1, Dnmt1, and Dnmt3a in SCNT embryos treated with Scriptaid were significantly lower than the control group at the eight-cell stage (p<0.05), but the expression of HAT1 and Dnmt1a was higher than the control group at the blastocyst stage (p<0.05). When 96 blastocysts developed from Scriptaid-treated SCNT embryos were transferred into 48 recipients, 11 recipients (22.9%) became pregnant, whereas only one recipient (11.1%) became pregnant following transfer of 18 blastocysts developed from untreated SCNT embryos into nine recipients. These results indicate that treatment of buffalo SCNT embryos with Scriptaid can improve their developmental competence, and this action is mediated by resulting in a similar histone acetylation level and global DNA methylation level compared to in vitro-fertilized embryos through regulating the expression pattern of genes related to histone acetylation and DNA methylation.

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Inhibition of histone deacetylases enhances DNA damage repair in SCNT embryos

Cited by 30 publications

References 68 publications

CDK2 Is Required for the DNA Damage Response During Porcine Early Embryonic Development

CDK2 Is Required for the DNA Damage Response During Porcine Early Embryonic Development

Double‐strand DNA breaks are mainly repaired by the homologous recombination pathway in early developing swine embryos

Effects of Scriptaid on the Histone Acetylation, DNA Methylation and Development of Buffalo Somatic Cell Nuclear Transfer Embryos

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