title: The HDAC Inhibitor LAQ824 Enhances Epigenetic Reprogramming and In Vitro Development of Porcine SCNT Embryos.
abstract: OBJECTIVE
  Hypoacetylation caused by aberrant epigenetic nuclear reprogramming results in low efficiency of mammalian somatic cell nuclear transfer (SCNT). Many epigenetic remodeling drugs have been used in attempts to improve in vitro development of porcine SCNT embryos. In this study, we examined the effects of LAQ824, a structurally novel histone acetylase inhibitor, on the nuclear reprogramming and in vitro development of porcine SCNT embryos.
  
  
  METHODS
  LAQ824 treatment was supplemented during the culture of SCNT embryos. The reprogramming levels were measured by immunofluorescence and quantified by image J software. Relative expression levels of 18 genes were analyzed by quantitative real-time PCR.
  
  
  RESULTS
  100 nM LAQ824 treatment of post-activation SCNT embryos for 24 h significantly improved the subsequent blastocyst formation rate. The LAQ824 treatment enhanced histone 3 lysine 9 (H3K9) levels, histone 4 lysine 12 (H4K12) levels, and reduced global DNA methylation levels as well as anti-5-methylcytosine (5-mC) at the pseudo-pronuclear and 2-cell stages. Furthermore, LAQ824 treatment positively regulated the mRNA expression of genes for histone acetylation (HAT1, HDAC1, 2, 3, and 6), DNA methylation (DNMT1, 3a and 3b), development (Pou5f1, Nanog, Sox2, and GLUT1) and apoptosis (Bax, Bcl2, Caspase 3 and Bak) in blastocysts.
  
  
  CONCLUSIONS
  Optimum exposure (100 nM for 24 h) to LAQ824 post-activation improved the in vitro development of porcine SCNT embryos by enhancing levels of H3K9 and H4K12, reducing 5-mC, and regulating gene expression.
authors.forename: Jun-Xue
authors.initials: JX
authors.lastname: Jin
authors.forename: Sanghoon
authors.initials: S
authors.lastname: Lee
authors.forename: Anukul
authors.initials: A
authors.lastname: Taweechaipaisankul
authors.forename: Geon A
authors.initials: GA
authors.lastname: Kim
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism; D000107:Acetylation / Q000187:drug effects; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D065150:Cellular Reprogramming / Q000187:drug effects*; D019175:DNA Methylation / Q000187:drug effects; D004622:Embryo, Mammalian; D015870:Gene Expression; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D013552:Swine
chemical_list: D015415:Biomarkers; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; C477361:LAQ824; D044503:5-Methylcytosine
keywords: In vitro development; Pig, Histone deacetylase inhibitor; Reprogramming; Somatic cell nuclear transfer