title: Beneficial effects of diazepin-quinazolin-amine derivative (BIX-01294) on preimplantation development and molecular characteristics of cloned mouse embryos.
abstract: Somatic cell nuclear transfer is frequently associated with abnormal epigenetic modifications that may lead to the developmental failure of cloned embryos. BIX-01294 (a diazepine-quinazoline-amine derivative) is a specific inhibitor of the histone methyltransferase G9a. The aim of the present study was to investigate the effects of BIX-01294 on development, dimethylation of histone H3 at lysine 9 (H3K9), DNA methylation and the expression of imprinted genes in cloned mouse preimplantation embryos. There were no significant differences in blastocyst rates of cloned embryos treated with or without 0.1μM BIX-01294. Relative to clone embryos treated without 0.1μM BIX-01294, exposure of embryos to BIX-01294 decreased histone H3K9 dimethylation and DNA methylation in cloned embryos to levels that were similar to those of in vivo-fertilised embryos at the 2-cell and blastocyst stages. Cloned embryos had lower expression of octamer-binding transcription factor 4 (Oct4) and small nuclear ribonucleoprotein N (Snrpn), but higher expression of imprinted maternally expressed transcript (non-protein coding) (H19) and growth factor receptor-bound protein 10 (Grb10) compared with in vivo-fertilised counterparts. The addition of 0.1μM BIX-01294 to the activation and culture medium resulted in lower H19 expression and higher cyclin dependent kinase inhibitor 1C (Cdkn1c) and delta-like 1 homolog (Dlk1) expression, but had no effect on the expression of Oct4, Snrpn and Grb10. The loss of methylation at the Grb10 cytosine-phosphorous-guanine (CpG) islands in cloned embryos was partially corrected by BIX-01294. These results indicate that BIX-01294 treatment of cloned embryos has beneficial effects in terms of correcting abnormal epigenetic modifications, but not on preimplantation development.
authors.affiliation: The First Affiliated Hospital, Fujian Medical University, Chazhong Road, 350005, Fuzhou, PR China.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Huang
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Jiang
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Miao
authors.initials: M
authors.lastname: Yu
authors.affiliation: The Second Affiliated Hospital, Fujian Medical University, Donghai Road, 362000, Quanzhou, PR China.
authors.forename: Rongfu
authors.initials: R
authors.lastname: Huang
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Yao
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Zheng
authors.affiliation: College of Preclinical Medicine, Fujian Medical University, Jiaotong Road, 350004, Fuzhou, PR China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Yang
mesh_terms: D000818:Animals; D001381:Azepines / Q000494:pharmacology*; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019976:Cloning, Organism / Q000662:veterinary*; D018899:CpG Islands / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects; D004477:Ectogenesis / Q000187:drug effects*; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005260:Female; D051383:GRB10 Adaptor Protein / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D011495:Histone-Lysine N-Methyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008297:Male; D008745:Methylation / Q000187:drug effects; D051379:Mice; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D010312:Parthenogenesis / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011799:Quinazolines / Q000494:pharmacology*
chemical_list: D001381:Azepines; C518299:BIX 01294; D004791:Enzyme Inhibitors; C495959:Grb10 protein, mouse; D006657:Histones; D011799:Quinazolines; D051383:GRB10 Adaptor Protein; C533711:G9a protein, mouse; D011495:Histone-Lysine N-Methyltransferase; D008239:Lysine