ArrayExpress Uploaderarrayexpress_sid458128 nMPfeifferDMSO (72h)EZH2 mutant28 nMPfeifferDMSO (72h)EZH2 mutant28 nMPfeifferGSK126 (500nM; 72h)EZH2 mutant28 nMPfeifferGSK126 (500nM; 72h)EZH2 mutant134 nM nMWSU-DLCL2DMSO (72h)EZH2 mutant134 nMWSU-DLCL2DMSO (72h)EZH2 mutant134 nMWSU-DLCL2GSK126 (500nM; 72h)EZH2 mutant134 nMWSU-DLCL2GSK126 (500nM; 72h)EZH2 mutant232 nMKARPAS-422DMSO (72h)EZH2 mutant232 nMKARPAS-422DMSO (72h)EZH2 mutant232 nMKARPAS-422GSK126 (500nM; 72h)EZH2 mutant232 nMKARPAS-422GSK126 (500nM; 72h)EZH2 mutant448 nMSU-DHL-10DMSO (72h)EZH2 mutant448 nMSU-DHL-10DMSO (72h)EZH2 mutant448 nMSU-DHL-10GSK126 (500nM; 72h)EZH2 mutant448 nMSU-DHL-10GSK126 (500nM; 72h)EZH2 mutant516 nMHtDMSO (72h)EZH2 wild type516 nMHtDMSO (72h)EZH2 wild type516 nMHtGSK126 (500nM; 72h)EZH2 wild type516 nMHtGSK126 (500nM; 72h)EZH2 wild type582 nMSU-DHL-6DMSO (72h)EZH2 mutant582 nMSU-DHL-6DMSO (72h)EZH2 mutant582 nMSU-DHL-6GSK126 (500nM; 72h)EZH2 mutant582 nMSU-DHL-6GSK126 (500nM; 72h)EZH2 mutant861 nMDBDMSO (72h)EZH2 mutant861 nMDBDMSO (72h)EZH2 mutant861 nMDBGSK126 (500nM; 72h)EZH2 mutant861 nMDBGSK126 (500nM; 72h)EZH2 mutant1019 nMOCI-LY-19DMSO (72h)EZH2 wild type1019 nMOCI-LY-19DMSO (72h)EZH2 wild type1019 nMOCI-LY-19GSK126 (500nM; 72h)EZH2 wild type1019 nMOCI-LY-19GSK126 (500nM; 72h)EZH2 wild typenot specifiedSU-DHL-4DMSO (72h)EZH2 mutantnot specifiedSU-DHL-4DMSO (72h)EZH2 mutantnot specifiedSU-DHL-4GSK126 (500nM; 72h)EZH2 mutantnot specifiedSU-DHL-4GSK126 (500nM; 72h)EZH2 mutant13786 nMToledoDMSO (72h)EZH2 wild type13786 nMToledoDMSO (72h)EZH2 wild type13786 nMToledoGSK126 (500nM; 72h)EZH2 wild type13786 nMToledoGSK126 (500nM; 72h)EZH2 wild type04We studied transcriptional changes by Affymetrix human microarrays in DLBCL cell lines as a result of treatment with GSK126, a potent,...23051747/profile/8773/arrayexpressuploader440b-cell lymphomacellchromatindiffuse large b-cell lymphomafollicular lymphomahistonehistone h3lymphomalysineDec.12, 2014Falsegene-expression-profiling-of-ezh2-mutant-and-wildhttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-40971E-GEOD-40971_A-AFFY-44Gene expression profiling of EZH2 mutant and wild type DLBCL cell lines treated with EZH2 inhibitorSep.19, 2014We studied transcriptional changes by Affymetrix human microarrays in DLBCL cell lines as a result of treatment with GSK126, a potent, highly-selective, SAM-competitive, small molecule inhibitor of EZH2 In eukaryotes, epigenetic 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 responsible for repressing target gene expression through methylation of histone H3 on lysine 27 (H3K27). Over-expression of EZH2 is implicated in tumorigenesis and correlates with poor prognosis in multiple tumor types. Recent reports have identified somatic heterozygous mutations of Y641 and A677 residues within the catalytic SET domain of EZH2 in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL). The Y641 residue is the most frequently mutated residue, with 22% of GCB (Germinal Cell B-cell) DLBCL and FL harboring mutations at this site. These lymphomas exhibit increased H3K27 tri-methylation (H3K27me3) due to altered substrate preferences of the mutant enzymes. However, it is unknown whether direct inhibition of EZH2 methyltransferase activity alone will be effective in treating lymphomas carrying activating EZH2 mutations. Herein, we demonstrate that GSK126, a potent, highly-selective, SAM-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 dramatically 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. 10 DLBCL cell lines (7 mutant and 3 wild type EZH2), that were differentially sensitive to GSK126 in proliferation assays, were treated for 72 hours, in duplicate (n=2), with either DMSO (vehicle) or 500nM of GSK126, a potent selective EZH2 inhibitor. EZH2 mutant cell lines are Pfeiffer, KARPAS-422, WSU-DLCL2, SU-DHL-10, SU-DHL-6, DB and SU-DHL-4. EZH2 wildtype cell lines are HT, OCI-LY-19 and Toledo.E-GEOD-40971humanhttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-40971/samples/