26 HISTONE DEACETYLASE INHIBITOR SCRIPTAID IMPROVES EPIGENETIC REPROGRAMMING AND CLONING EFFICIENCY IN THE PIG

2015 ◽  
Vol 27 (1) ◽  
pp. 105
Author(s):  
S. Liang ◽  
T. Kim ◽  
N.-H. Kim ◽  
X.-S. Cui
Keyword(s):  
Histone Deacetylase ◽  
Histone Deacetylase Inhibitor ◽  
Histone H3 ◽  
Development Program ◽  
Donor Cell ◽  
Epigenetic Reprogramming ◽  
Republic Of Korea ◽  
Cloning Efficiency ◽  
Cell Stage ◽  
Deacetylase Inhibitor

After somatic cell nuclear transfer (SCNT), the epigenetic state of a differentiated donor cell nucleus must be reversed to the embryonic state. Incomplete epigenetic reprogramming and abnormal gene activation of the donor cell nuclei is thought to be the cause of low cloning efficiency. To improve cloning efficiency, we investigated the effect of scriptaid, a novel histone deacetylase inhibitor, on the in vitro development of porcine SCNT embryos were investigated. Cumulus cells collected from cumulus-oocyte complexes (COC) after 44 h of maturation were used for donor cell, and embryos were cultured in porcine zygote medium (PZM)-5 medium for 7 days. We found that treating SCNT embryos with 300 or 500 nM scriptaid for 20 h after activation increased developmental rate to the blastocyst stage (300 nM, 26.2%; 500 nM, 24.6% v. 100 nM, 18.3%; Ctrl, 15.7%; P < 0.05) and total cell numbers (300 nM, 43.5; 500 nM, 40.8 v. 100 nM, 33.8; Ctrl, 32.3; P < 0.05). Additionally, results of the TUNEL assay indicated that scriptaid decreased apoptosis (300 nM, 6.8% v. Ctrl, 11.4%; P < 0.05) in SCNT blastocysts. After the 300 nM scriptaid treatment, the levels of acetylated histone H3 lysine 9 and 5-hydroxymethylcytosines were increased (P < 0.05), and histone H3 lysine 9 trimethylation and 5-methylcytosine were decreased at the 1-cell stage, which might explain the enhanced (P < 0.05) transcript levels of mir-152, Oct4, Cdx2, and Bcl-xL and reduced (P < 0.05) transcription of Dnmt1, Casp3, and Bax in blastocysts. In conclusion, scriptaid enhances the developmental capacity by preventing apoptosis, and improves nuclear reprogramming in porcine SCNT embryos.This work was supported by the Bio-industry Technology Development Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea, and by a grant from the Next-Generation BioGreen 21 Program (No. PJ009601 and PJ009098), Rural Development Administration, Republic of Korea.

Histone deacetylase inhibitor FR901228 enhances adenovirus infection of hematopoietic cells

Blood ◽  
2002 ◽  
Vol 99 (6) ◽  
pp. 2248-2251 ◽  
Author(s):  
Masaki Kitazono ◽  
Vemulkonda Koneti Rao ◽  
Rob Robey ◽  
Takashi Aikou ◽  
Susan Bates ◽  
...  
Keyword(s):  
Histone Deacetylase ◽  
Peripheral Blood ◽  
Histone Deacetylase Inhibitor ◽  
Mononuclear Cells ◽  
Histone H3 ◽  
Hematopoietic Cells ◽  
Adenovirus Infection ◽  
Peripheral Blood Stem Cells ◽  
Peripheral Blood Mononuclear ◽  
Deacetylase Inhibitor

Abstract Adenovirus infection of hematopoietic cells frequently requires high virus concentrations and long incubation times to obtain moderate infection levels because these cells have low levels of Coxsackie and adenovirus receptor (CAR) and αv integrin. The effect of treatment with FR901228 (depsipeptide), a histone deacetylase inhibitor in phase 2 clinical trials, was studied in K562 cells, granulocyte–colony-stimulating factor–mobilized peripheral blood mononuclear cells (PBMCs), and CD34+ peripheral blood stem cells (PBSCs). FR901228 increased CAR and αvintegrin RNA levels and histone H3 acetylation. FR901228 treatment before adenovirus infection was associated with at least a 10-fold increase in transgene expression from a β-galactosidase–expressing adenoviral vector. More than 80% of the PBMCs or CD34+ PBSCs from 7 different donors were β-galactosidase–positive after adenovirus infection with a multiplicity of infection of 10 for 60 minutes. Increased CAR, αv integrin, and acetylated histone H3 levels were observed in PBMCs from a patient treated with FR901228. These studies suggest that FR901228 can increase the efficiency of adenoviral infection in hematopoietic cells.

scholarly journals Acetylation of Histone H3 and Adrenergic-Regulated Gene Transcription in Rat Pinealocytes

Endocrinology ◽  
2007 ◽  
Vol 148 (10) ◽  
pp. 4592-4600 ◽  
Author(s):  
A. K. Ho ◽  
D. M. Price ◽  
W. G. Dukewich ◽  
N. Steinberg ◽  
T. G. Arnason ◽  
...  
Keyword(s):  
Histone Acetylation ◽  
Histone Deacetylase ◽  
Gene Transcription ◽  
Histone Deacetylase Inhibitor ◽  
Trichostatin A ◽  
Histone H3 ◽  
Dna Recovery ◽  
Inducible Camp Early Repressor ◽  
Deacetylase Inhibitor ◽  
Induced Genes

In this study we investigated the effect of histone acetylation on the transcription of adrenergic-induced genes in rat pinealocytes. We found that treatment of pinealocytes with trichostatin A (TSA), a histone deacetylase inhibitor, caused hyperacetylation of histone H3 (H3) Lys14 at nanomolar concentrations. Hyperacetylation of H3 was also observed after treatment with scriptaid, a structurally unrelated histone deacetylase inhibitor. The effects of TSA and scriptaid were inhibitory on the adrenergic induction of arylalkylamine-n-acetyltransferase (aa-nat) mRNA, protein, and enzyme activity, and on melatonin production. TSA at higher concentrations also inhibited the adrenergic induction of mapk phosphatase-1 (mkp-1) and inducible cAMP early repressor mRNAs. In contrast, the effect of TSA on the norepinephrine induction of the c-fos mRNA was stimulatory. Moreover, the effect of TSA on adrenergic-induced gene transcription was dependent on the time of its addition; its effect was only observed during the active phase of transcription. Chromatin immunoprecipitation with antibodies against acetylated Lys14 of H3 showed an increase in DNA recovery of the promoter regions of aa-nat, mkp-1, and c-fos after treatment with TSA. Together, our results demonstrate that histone acetylation differentially influences the transcription of adrenergic-induced genes, an enhancing effect for c-fos but inhibitory for aa-nat, mkp-1, and inducible cAMP early repressor. Moreover, both inhibitory and enhancing effects appear to be mediated through specific modification of promoter-bound histones during gene transcription.

41 DEVELOPMENT OF SOMATIC CELL NUCLEAR TRANSFERRED RAT EMBRYOS TO THE BLASTOCYST BY A HISTONE DEACETYLASE INHIBITOR, TRICHOSTATIN A

2011 ◽  
Vol 23 (1) ◽  
pp. 126
Author(s):  
N. Kashiwazaki ◽  
N. Nakajima ◽  
K. Fujimaki ◽  
K. Syudo ◽  
J. Ito
Keyword(s):  
Somatic Cell ◽  
Histone Deacetylase ◽  
Histone Deacetylase Inhibitor ◽  
Trichostatin A ◽  
Cell Stage ◽  
Rat Embryos ◽  
Reconstructed Embryos ◽  
Deacetylase Inhibitor ◽  
Histone Deacetylase Inhibitor Trichostatin ◽  
Pronuclear Formation

Although cloned animals have been reproduced in many species via somatic cell nuclear transfer (SCNT), only 1 previous report showed successful generation of cloned rats. Some researchers evaluated developmental competence of reconstructed rat embryos and showed that these reconstructed embryos were never progressed beyond the 2-cell stage because of inadequate formation of the mitotic assembly and nuclear organisation or improper transcription of cytoskeleton genes. Even though our group also improved the proportion of 2 pronuclear formation in rat reconstructed embryos by the treatment with a proteasome inhibitor, MG132 (Nakajima et al. 2008 Cloning and Stem Cells 10, 461–468), no embryos were developed beyond the 2-cell stage. Recently, it has been demonstrated that a histone deacetylase inhibitor, trichostatin A (TSA) treatment dramatically improves efficiency of cloning in the mouse, although the precise effect of TSA is unclear in SCNT. However, in the case of rats, availability of TSA has not been tested. The aim of the present study was to clarify whether TSA treatment was also effective in rat SCNT. According to our previous reports, rat oocytes were collected and the spindles of the oocytes were removed in medium supplemented with MG132. After enucleation, nuclei of cumulus cells were injected in the oocytes and the reconstructed embryos were cultured in the medium supplemented with 0 (control), 5, 50, or 500 nM TSA. More than 100 embryos are used for each treatment group. Pronuclear formation, development to the 2-cell stage and blastocyst formation were evaluated. Our results demonstrated that TSA treatment affected neither the proportion of pronuclear formation nor development to the 2-cell stage in rat reconstructed embryos. However, the treatment with higher concentrations of TSA treatment (50 nM and 500 nM) compared to the concentration (5 nM) which was usually used for mouse SCNT enabled the reconstructed embryos to develop to the blastocyst stage but at low rate (1.4 to 2.2%). In both the 5 nM treatment and control groups, none of the rat reconstructed embryos developed to blastocyst. Taken together, our data suggests that TSA treatment also seems to be effective in rat SCNT. These findings will be useful for improvement of the protocol in rat SCNT. This work was also supported in part by the Promotion and Mutual Aid Corporation for Private Schools of Japan, Grant-in-Aid for Matching Fund Subsidy for Private Universities to J. I. and N. K.

scholarly journals Combined epigenetic therapy with the histone methyltransferase EZH2 inhibitor 3-deazaneplanocin A and the histone deacetylase inhibitor panobinostat against human AML cells

Blood ◽  
2009 ◽  
Vol 114 (13) ◽  
pp. 2733-2743 ◽  
Author(s):  
Warren Fiskus ◽  
Yongchao Wang ◽  
Arun Sreekumar ◽  
Kathleen M. Buckley ◽  
Huidong Shi ◽  
...  
Keyword(s):  
Histone Deacetylase ◽  
Histone Deacetylase Inhibitor ◽  
Hox Genes ◽  
Severe Combined Immunodeficiency ◽  
Histone H3 ◽  
Histone Methyltransferase ◽  
Epigenetic Therapy ◽  
Core Proteins ◽  
Deacetylase Inhibitor ◽  
Induced Apoptosis

Abstract The polycomb repressive complex (PRC) 2 contains 3 core proteins, EZH2, SUZ12, and EED, in which the SET (suppressor of variegation–enhancer of zeste-trithorax) domain of EZH2 mediates the histone methyltransferase activity. This induces trimethylation of lysine 27 on histone H3, regulates the expression of HOX genes, and promotes proliferation and aggressiveness of neoplastic cells. In this study, we demonstrate that treatment with the S-adenosylhomocysteine hydrolase inhibitor 3-deazaneplanocin A (DZNep) depletes EZH2 levels, and inhibits trimethylation of lysine 27 on histone H3 in the cultured human acute myeloid leukemia (AML) HL-60 and OCI-AML3 cells and in primary AML cells. DZNep treatment induced p16, p21, p27, and FBXO32 while depleting cyclin E and HOXA9 levels. Similar findings were observed after treatment with small interfering RNA to EZH2. In addition, DZNep treatment induced apoptosis in cultured and primary AML cells. Furthermore, compared with treatment with each agent alone, cotreatment with DZNep and the pan-histone deacetylase inhibitor panobinostat caused more depletion of EZH2, induced more apoptosis of AML, but not normal CD34+ bone marrow progenitor cells, and significantly improved survival of nonobese diabetic/severe combined immunodeficiency mice with HL-60 leukemia. These findings indicate that the combination of DZNep and panobinostat is effective and relatively selective epigenetic therapy against AML cells.

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