IVMBIX-01294, an inhibitor of the histone methyltransferase EHMT2, disrupts histone H3 lysine 9 (H3K9) dimethylation in the cleavage-stage porcine embryo.
Global patterns of histone methylation are remodelled during cleavage development. Of the five histone methyltransferases known to mediate methylation of the lysine 9 residue of histone H3 (H3K9), euchromatic histone-lysine N-methyltransferase 2 (EHMT2; also known as G9a) has been shown to be a primary mediator of H3K9 dimethylation; BIX-01294 has been shown to be a specific inhibitor of EHMT2. The objective of the present study was to determine the effect of BIX-01294 treatment on global H3K9 dimethylation in porcine embryos. We hypothesised that inhibition of EHMT2 by BIX-01294 would result in reduced levels of H3K9 dimethylation and compromised embryo development. Our results showed that incubation in 5µM BIX-01294 markedly reduced global levels of H3K9 dimethylation at the pronuclear, 2-cell and 4-cell stages of development and resulted in developmental arrest before blastocyst formation. Although transient exposure of embryos to BIX-01294 did not alter in vitro development, embryos transiently exposed to BIX-01294 did not establish pregnancy. These data demonstrate that BIX-01294 is a potent inhibitor of H3K9 dimethylation and that transient alterations in global histone modifications can have profound effects on embryo developmental potential.
Department of Animal Sciences, Purdue University, 915 West State Street, West Lafayette, IN 47907, USA.
Ki-Eun
KE
Park
Christine M
CM
Johnson
Ryan A
RA
Cabot
D000818:Animals; D001381:Azepines / Q000494:pharmacology*; D002970:Cleavage Stage, Ovum / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000201:enzymology; D047108:Embryonic Development / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D011495:Histone-Lysine N-Methyltransferase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D008239:Lysine; D008745:Methylation; D011247:Pregnancy; D018873:Pregnancy Rate; D011799:Quinazolines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
D001381:Azepines; C518299:BIX 01294; D004791:Enzyme Inhibitors; D006657:Histones; D011799:Quinazolines; D012333:RNA, Messenger; D011495:Histone-Lysine N-Methyltransferase; D008239:Lysine