Identification of novel isoform-selective inhibitors within class I histone deacetylases.
Histone deacetylases (HDACs) represent an expanding family of protein modifying-enzymes that play important roles in cell proliferation, chromosome remodeling, and gene transcription. We have previously shown that recombinant human HDAC8 can be expressed in bacteria and retain its catalytic activity. To further explore the catalytic activity of HDACs, we expressed two additional human class I HDACs, HDAC1 and HDAC3, in baculovirus. Recombinant HDAC1 and HDAC3 fusion proteins remained soluble and catalytically active and were purified to near homogeneity. Interestingly, trichostatin (TSA) was found to be a potent inhibitor for all three HDACs (IC50 value of approximately 0.1-0.3 microM), whereas another HDAC inhibitor MS-27-275 (N-(2-aminophenyl)-4-[N-(pyridin-3-methyloxycarbonyl)-aminomethyl]benzamide) preferentially inhibited HDAC1 (IC50 value of approximately 0.3 microM) versus HDAC3 (IC50 value of approximately 8 microM) and had no inhibitory activity toward HDAC8 (IC50 value >100 microM). MS-27-275 as well as TSA increased histone H4 acetylation, induced apoptosis in the human colon cancer cell line SW620, and activated the simian virus 40 early promoter. HDAC1 protein was more abundantly expressed in SW620 cells compared with that of HDAC3 and HDAC8. Using purified recombinant HDAC proteins, we identified several novel HDAC inhibitors that preferentially inhibit HDAC1 or HDAC8. These inhibitors displayed distinct properties in inducing histone acetylation and reporter gene expression. These results suggest selective HDAC inhibitors could be identified using recombinantly expressed HDACs and that HDAC1 may be a promising therapeutic target for designing HDAC inhibitors for proliferative diseases such as cancer.
Hu
Erding
E
Department of Vascular Biology, GlaxoSmithKline, Rm. 25-1084, 709 Swedeland Road, King of Prussia, PA 19406, USA. erding_hu-1@gsk.com
Dul
Edward
E
Sung
Chiu-Mei
CM
Chen
Zunxuan
Z
Kirkpatrick
Robert
R
Zhang
Gui-Feng
GF
Johanson
Kyung
K
Liu
Ronggang
R
Lago
Amparo
A
Hofmann
Glenn
G
Macarron
Ricardo
R
de los Frailes
Maite
M
Perez
Paloma
P
Krawiec
John
J
Winkler
James
J
Jaye
Michael
M
D001549:Benzamides / Q000494:pharmacology*; D003001:Cloning, Molecular; D003110:Colonic Neoplasms / Q000473:pathology; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression; D056284:Histone Deacetylase 1; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D020033:Protein Isoforms / Q000037:antagonists & inhibitors; D011725:Pyridines / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D000099186:Histone Deacetylase 3
D001549:Benzamides; D004791:Enzyme Inhibitors; D056284:Histone Deacetylase 1; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006877:Hydroxamic Acids; D020033:Protein Isoforms; D011725:Pyridines; D011994:Recombinant Proteins; D000099186:Histone Deacetylase 3; C118739:entinostat; C012589:trichostatin A; C110550:HDAC1 protein, human