Abstract
In eukaryotes, post-translational modification of histones is critical for regulation of chromatin structure and gene expression. EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and is involved in repressing gene expression through methylation of histone H3 on lysine 27 (H3K27). EZH2 overexpression is implicated in tumorigenesis and correlates with poor prognosis in several tumour types. Additionally, somatic heterozygous mutations of Y641 and A677 residues within the catalytic SET domain of EZH2 occur in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma. The Y641 residue is the most frequently mutated residue, with up to 22% of germinal centre B-cell DLBCL and follicular lymphoma harbouring mutations at this site. These lymphomas have increased H3K27 tri-methylation (H3K27me3) owing to altered substrate preferences of the mutant enzymes. However, it is unknown whether specific, direct inhibition of EZH2 methyltransferase activity will be effective in treating EZH2 mutant lymphomas. Here we demonstrate that GSK126, a potent, highly selective, S-adenosyl-methionine-competitive, small-molecule inhibitor of EZH2 methyltransferase activity, decreases global H3K27me3 levels and reactivates silenced PRC2 target genes. GSK126 effectively inhibits the proliferation of EZH2 mutant DLBCL cell lines and markedly inhibits the growth of EZH2 mutant DLBCL xenografts in mice. Together, these data demonstrate that pharmacological inhibition of EZH2 activity may provide a promising treatment for EZH2 mutant lymphoma.
MeSH Terms
Animals
Cell Line, Tumor
Cell Proliferation/drug effects
Enhancer of Zeste Homolog 2 Protein
Gene Expression Regulation, Neoplastic/drug effects
Gene Silencing/drug effects
Histone Methyltransferases
Histone-Lysine N-Methyltransferase/antagonists & inhibitors,genetics,metabolism
Histones/chemistry,metabolism
Humans
Indoles/pharmacology,therapeutic use
Lymphoma, Follicular/drug therapy,enzymology,genetics,pathology
Lymphoma, Large B-Cell, Diffuse/drug therapy,enzymology,genetics,pathology
Methylation/drug effects
Mice
Mutation/genetics
Neoplasm Transplantation
Polycomb Repressive Complex 2/antagonists & inhibitors,genetics,metabolism
Pyridones/pharmacology,therapeutic use
Repressor Proteins/chemistry,metabolism
Transcriptional Activation/drug effects
Transplantation, Heterologous
Chemicals
GSK-2816126
Histones
Indoles
Pyridones
Repressor Proteins
Histone Methyltransferases
EZH2 protein, human
Enhancer of Zeste Homolog 2 Protein
Ezh2 protein, mouse
Histone-Lysine N-Methyltransferase
Polycomb Repressive Complex 2
Authors & Affiliations
22 authors, click to expand affiliations / ORCID
McCabe Michael T
Cancer Epigenetics Discovery Performance Unit, Cancer Research, Oncology R&D, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
Ott Heidi M
Ganji Gopinath
Korenchuk Susan
Thompson Christine
Van Aller Glenn S
Liu Yan
Graves Alan P
Della Pietra Anthony
Diaz Elsie
LaFrance Louis V
Mellinger Mark
Duquenne Celine
Tian Xinrong
Kruger Ryan G
McHugh Charles F
Brandt Martin
Miller William H
Dhanak Dashyant
Verma Sharad K
Tummino Peter J
Creasy Caretha L
References (30)
30 references, click to expand
-
EZH2 codon 641 mutations are common in BCL2-rearranged germinal center B cell lymphomas.
PLoS One. 2011;6(12):e28585
PMID: 22194861
-
Ultrafast and memory-efficient alignment of short DNA sequences to the human genome.
Genome Biol. 2009;10(3):R25
PMID: 19261174
-
Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma.
BMC Cancer. 2010 Oct 04;10:524
PMID: 20920340
-
The Y641C mutation of EZH2 alters substrate specificity for histone H3 lysine 27 methylation states.
FEBS Lett. 2011 Oct 3;585(19):3011-4
PMID: 21856302
-
Many paths to methyltransfer: a chronicle of convergence.
Trends Biochem Sci. 2003 Jun;28(6):329-35
PMID: 12826405
-
Coordinated activities of wild-type plus mutant EZH2 drive tumor-associated hypertrimethylation of lysine 27 on histone H3 (H3K27) in human B-cell lymphomas.
Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):20980-5
PMID: 21078963
-
Somatic mutations at EZH2 Y641 act dominantly through a mechanism of selectively altered PRC2 catalytic activity, to increase H3K27 trimethylation.
Blood. 2011 Feb 24;117(8):2451-9
PMID: 21190999
-
The roles of EZH2 in cell lineage commitment.
Am J Transl Res. 2011 May 15;3(3):243-50
PMID: 21654879
-
A clustering approach for identification of enriched domains from histone modification ChIP-Seq data.
Bioinformatics. 2009 Aug 1;25(15):1952-8
PMID: 19505939
-
Validation of the histone methyltransferase EZH2 as a therapeutic target for various types of human cancer and as a prognostic marker.
Cancer Sci. 2011 Jul;102(7):1298-305
PMID: 21539681
-
Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin.
Nat Genet. 2010 Feb;42(2):181-5
PMID: 20081860
-
Anaplastic lymphoma kinase inhibition in non-small-cell lung cancer.
N Engl J Med. 2010 Oct 28;363(18):1693-703
PMID: 20979469
-
Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders.
Nat Genet. 2010 Aug;42(8):722-6
PMID: 20601953
-
EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells.
Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11606-11
PMID: 14500907
-
Frequent mutation of histone-modifying genes in non-Hodgkin lymphoma.
Nature. 2011 Jul 27;476(7360):298-303
PMID: 21796119
-
Development and validation of reagents and assays for EZH2 peptide and nucleosome high-throughput screens.
J Biomol Screen. 2012 Dec;17(10):1279-92
PMID: 22904200
-
A new human B-cell non-Hodgkin's lymphoma cell line (Karpas 422) exhibiting both t (14;18) and t(4;11) chromosomal translocations.
Blood. 1990 Feb 1;75(3):709-14
PMID: 2297573
-
The polycomb group protein EZH2 is involved in progression of prostate cancer.
Nature. 2002 Oct 10;419(6907):624-9
PMID: 12374981
-
Improved survival with vemurafenib in melanoma with BRAF V600E mutation.
N Engl J Med. 2011 Jun 30;364(26):2507-16
PMID: 21639808
-
DZNep is a global histone methylation inhibitor that reactivates developmental genes not silenced by DNA methylation.
Mol Cancer Ther. 2009 Jun;8(6):1579-88
PMID: 19509260
-
Analysis of the coding genome of diffuse large B-cell lymphoma.
Nat Genet. 2011 Jul 31;43(9):830-7
PMID: 21804550
-
Mutation of A677 in histone methyltransferase EZH2 in human B-cell lymphoma promotes hypertrimethylation of histone H3 on lysine 27 (H3K27).
Proc Natl Acad Sci U S A. 2012 Feb 21;109(8):2989-94
PMID: 22323599
-
Somatic mutations of the histone H3K27 demethylase gene UTX in human cancer.
Nat Genet. 2009 May;41(5):521-3
PMID: 19330029
-
Mutational spectrum analysis of chronic myelomonocytic leukemia includes genes associated with epigenetic regulation: UTX, EZH2, and DNMT3A.
Blood. 2011 Oct 6;118(14):3932-41
PMID: 21828135
-
Novel homo- and hemizygous mutations in EZH2 in myeloid malignancies.
Leukemia. 2010 Oct;24(10):1799-804
PMID: 20724984
-
Bryostatin 1 down-regulates mdr1 and potentiates vincristine cytotoxicity in diffuse large cell lymphoma xenografts.
Clin Cancer Res. 1998 May;4(5):1305-14
PMID: 9607591
-
Kinetic applications using high substrate and competitive inhibitor concentrations to determine Ki or Km.
Anal Biochem. 1994 Aug 15;221(1):53-6
PMID: 7985803
-
Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction.
Biochem Pharmacol. 1973 Dec 1;22(23):3099-108
PMID: 4202581
-
Genomic loss of microRNA-101 leads to overexpression of histone methyltransferase EZH2 in cancer.
Science. 2008 Dec 12;322(5908):1695-9
PMID: 19008416
-
Frequent mutations of chromatin remodeling genes in transitional cell carcinoma of the bladder.
Nat Genet. 2011 Aug 07;43(9):875-8
PMID: 21822268