Effects of the histone deacetylase inhibitor ‘Scriptaid’ on the developmental competence of mouse embryos generated through round spermatid injection

Kong, Pengcheng; Yin, Minzhi; Chen, Dongbao; Li, Shangang; Li, Yao; Xing, Fengying; Jiang, Man-Xi; Fang, Zhenfu; Lyu, Qifeng; Chen, Xuejin

doi:10.1093/humrep/dew290

Cited by 3 publications

(10 citation statements)

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“…Thus far, various strategies of epigenomic modulation (epigenetic transformation) that are mediated by non-specific inhibitors of histone deacetylases (HDACi) and/or DNA methyltransferases (DNMTi) have been applied to improve reprogrammability of donor cell-inherited genome in somatic cell nuclear transfer (SCNT)-derived oocytes and resultant cloned embryos in different mammalian species [12][13][14][15][16]. Analogous methods of HDACiand/or DNMTi-dependent epigenetic transformation have been adapted to enhance capabilities of parental genomes to be epigenetically reprogrammed in the in vitro fertilization (IVF)-derived embryos [17][18][19][20]. Such approaches have also been used either to expedite the epigenomic reprogramming and molecular dedifferentiation of adult somatic cells or mesenchymal stem cells (MSCs) into induced pluripotent stem cells (iPSCs) [21][22][23][24][25] or to facilitate the molecular rejuvenation of adult MSCs [21,26,27].…”

Section: Introductionmentioning

confidence: 99%

Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

Wiater

Samiec

Skrzyszowska

et al. 2021

IJMS

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This study was conducted to explore whether trichostatin A-assisted epigenomic modulation (TSA-EM) can affect the expression of not only recombinant human α1,2-fucosyltransferase (rhα1,2-FT) and α-galactosidase A (rhα-Gal A) immune system enzymes but also Galα1→3Gal epitopes in ex vivo proliferating adult cutaneous fibroblast cells (ACFCs) derived from hFUT2×hGLA bi-transgenic pigs that had been produced for the needs of future xenotransplantation efforts. The ACFC lines were treated with 50 nM TSA for 24 h and then the expression profiles of rhα1,2-FT and rhα-Gal A enzymes were analyzed by Western blot and immunofluorescence. The expression profiles of the Galα1→3Gal epitope were determined by lectin blotting and lectin fluorescence. The ACFCs derived from non-transgenic (nTG) pigs were served as the negative (TSA−) and positive (TSA+) control groups. For both hFUT2×hGLA and nTG samples, the expression levels of α1,2-FT and α-Gal A proteins in TSA+ cells were more than twofold higher in comparison to TSA− cells. Moreover, a much lower expression of the Galα1→3Gal epitopes was shown in TSA− hFUT2×hGLA cells as compared to the TSA− nTG group. Interestingly, the levels of Galα1→3Gal expression in TSA-treated hFUT2×hGLA and nTG ACFCs were significantly higher than those noticed for their TSA-untreated counterparts. Summing up, ex vivo protection of effectively selected bi-transgenic ACFC lines, in which TSA-dependent epigenetic transformation triggered the enhancements in reprogrammability and subsequent expression of hFUT2 and hGLA transgenes and their corresponding transcripts, allows for cryopreservation of nuclear donor cells, nuclear-transferred female gametes, and resultant porcine cloned embryos. The latter can be used as a cryogenically conserved genetic resource of biological materials suitable for generation of bi-transgenic cloned offspring in pigs that is targeted at biomedical research in the field of cell/tissue xenotransplantation.

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Section: Introductionmentioning

confidence: 99%

Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

Wiater

Samiec

Skrzyszowska

et al. 2021

IJMS

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“…Mouse oocyte collection process was shown in our previous study [ 14 , 15 ]. Female C57BL/6 mice (4–6 weeks old) were used for superovulation.…”

Section: Methodsmentioning

confidence: 99%

“…Bisulfite mutagenesis was performed as previously described [ 15 ], with one blastocyst in each pool. Briefly, the DNA was wrapped in agarose gels to form little beads.…”

Section: Methodsmentioning

confidence: 99%

“…Methylation-specific PCR (MS-PCR) primers were used to match the allele-specific DNA strands (Supplementary Table S1). Following nested PCR, cloning and sequencing were performed as previously described [ 15 ]. Briefly, the first round of PCR was conducted using one bead containing bisulfite-treated DNA with the outer primers and Taq HS premix (TaKaRa).…”

Section: Methodsmentioning

confidence: 99%

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DNA methylation and gene expression changes in mouse pre- and post-implantation embryos generated by intracytoplasmic sperm injection with artificial oocyte activation

Yin

et al. 2021

Reprod Biol Endocrinol

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Background The application of artificial oocyte activation (AOA) after intracytoplasmic sperm injection (ICSI) is successful in mitigating fertilization failure problems in assisted reproductive technology (ART). Nevertheless, there is no relevant study to investigate whether AOA procedures increase developmental risk by disturbing subsequent gene expression at different embryonic development stages. Methods We used a mouse model to explore the influence of AOA treatment on pre- and post-implantation events. Firstly, the developmental potential of embryos with or without AOA treatment were assessed by the rates of fertilization and blastocyst formation. Secondly, transcriptome high-throughput sequencing was performed among the three groups (ICSI, ICSI-AOA and dICSI-AOA groups). The hierarchical clustering and Principal Component Analysis (PCA) analysis were used. Subsequently, Igf2r/Airn methylation analysis were detected using methylation-specific PCR sequencing following bisulfite treatment. Finally, birth rate and birth weight were examined following mouse embryo transfer. Results The rates of fertilization and blastocyst formation were significantly lower in oocyte activation-deficient sperm injection group (dICSI group) when compared with the ICSI group (30.8 % vs. 84.4 %, 10.0 % vs. 41.5 %). There were 133 differentially expressed genes (DEGs) between the ICSI-AOA group and ICSI group, and 266 DEGs between the dICSI-AOA group and ICSI group. In addition, the imprinted gene, Igf2r is up regulated in AOA treatment group compared to control group. The Igf2r/Airn imprinted expression model demonstrates that AOA treatment stimulates maternal allele-specific mehtylation spreads at differentially methylated region 2, followed by the initiation of paternal imprinted Airn long non-coding (lnc) RNA, resulting in the up regulated expression of Igf2r. Furthermore, the birth weight of newborn mice originating from AOA group was significantly lower compared to that of ICSI group. The pups born following AOA treatment did not show any other abnormalities during early development. All offspring mated successfully with fertile controls. Conclusions AOA treatment affects imprinted gene Igf2r expression and mehtylation states in mouse pre- and post-implantation embryo, which is regulated by the imprinted Airn. Nevertheless, no significant differences were found in post-natal growth of the pups in the present study. It is hoped that this study could provide valuable insights of AOA technology in assisted reproduction biology.

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“…Our studies have elucidated the main cause of poor clinical outcome as derived from epigenetic abnormalities by differences of nuclear protein in round spermatid (histone) and mature spermatozoa (protamine). 21 , 22 , 23 …”

Section: Introductionmentioning

confidence: 99%

How to improve the clinical outcome of round spermatid injection (ROSI) into the oocyte: Correction of epigenetic abnormalities

Tanaka

Watanabe

2023

Reprod Medicine & Biology

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Background First successful human round spermatid injection (ROSI) was conducted by Tesarik et al. in 1996 for the sole treatment of nonobstructive azoospermic men whose most advanced spermatogenic cells were elongating round spermatids. Nine offsprings from ROSI were reported between 1996 and 2000. No successful deliveries were reported for 15 years after that. Tanaka et al. reported 90 babies born after ROSI and their follow‐up studies in 2015 and 2018 showed no significant differences in comparison with those born after natural conception in terms of physical and cognitive abilities. However, clinical outcomes remain low. Method Clinical and laboratory data of successful cases in the precursor ROSI groups and those of Tanaka et al. were reviewed. Results Differences were found between the two groups in terms of identification of characteristics of round spermatid and oocyte activation. Additionally, epigenetic abnormalities were identified as underlying causes for poor ROSI results, besides correct identification of round spermatid and adequate oocyte activation. Correction of epigenetic errors could lead to optimal embryonic development. Conclusion Correction of epigenetic abnormalities has a probability to improve the clinical outcome of ROSI.

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Effects of the histone deacetylase inhibitor ‘Scriptaid’ on the developmental competence of mouse embryos generated through round spermatid injection

Cited by 3 publications

References 46 publications

Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

DNA methylation and gene expression changes in mouse pre- and post-implantation embryos generated by intracytoplasmic sperm injection with artificial oocyte activation

How to improve the clinical outcome of round spermatid injection (ROSI) into the oocyte: Correction of epigenetic abnormalities

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