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Active MYC signaling confers susceptibility to small-molecule SUMO inhibition in diffuse large B-cell lymphoma cell lines. (A) Hierarchical clustering (Euclidean/Ward) of indicated SUMO core components of normalized human diffuse large B-cell lymphoma (DLBCL) transcriptome profiles (GSE98588) revealed SUMO high and SUMO low subgroups as indicated. (B) Gene set enrichment analysis of SUMO high and SUMO low subgroups using Hallmark gene sets from the molecular-signature database (MSigDb). Indicated MYC target gene signatures have been identified using GeneTrail v3 (Kolgomorov-Smirnov-Test). Adjusted P values (q) are indicated. (C) Flow cytometry analysis of relative viability of indicated DLBCL cell lines treated with increasing SUMO inhibitor <t>(SUMOi)</t> <t>concentrations</t> <t>(Tak-981,</t> 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 hours (h) (n=3). (D) Gene set enrichment analysis of SUMOi-responder (Oci-Ly19, SU-DHL-5, SU-DHL-8) vs . SUMOi-non-responder (U-2932, SUDHL-4, DB, NU-DHL-1) on expression profiles accessed via GSE53798. Enrichment plot on SUMOylation signatures, obtained from the Reactome knowledgebase. Adjusted P values (q) are indicated. Volcano plot displays significant gene signatures of the Hallmark gene set (MSigDb) with both MYC Hallmark signatures (V1, V2) highlighted. (E) Enrichment plot on MYC Hallmark signatures, based on the analysis described in (D). Adjusted P values (q) are indicated. (F) Relative viability of Oci-Ly1 cells transduced with a MYC expression plasmid or a control plasmid, treated with increasing SUMOi concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 h (n=3). P values refers to 2,000 nM SUMOi and is determined by unpaired t -test. Immunoblot analysis of Oci-Ly1 control and MYC cell lines.
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Active MYC signaling confers susceptibility to small-molecule SUMO inhibition in diffuse large B-cell lymphoma cell lines. (A) Hierarchical clustering (Euclidean/Ward) of indicated SUMO core components of normalized human diffuse large B-cell lymphoma (DLBCL) transcriptome profiles (GSE98588) revealed SUMO high and SUMO low subgroups as indicated. (B) Gene set enrichment analysis of SUMO high and SUMO low subgroups using Hallmark gene sets from the molecular-signature database (MSigDb). Indicated MYC target gene signatures have been identified using GeneTrail v3 (Kolgomorov-Smirnov-Test). Adjusted P values (q) are indicated. (C) Flow cytometry analysis of relative viability of indicated DLBCL cell lines treated with increasing SUMO inhibitor <t>(SUMOi)</t> <t>concentrations</t> <t>(Tak-981,</t> 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 hours (h) (n=3). (D) Gene set enrichment analysis of SUMOi-responder (Oci-Ly19, SU-DHL-5, SU-DHL-8) vs . SUMOi-non-responder (U-2932, SUDHL-4, DB, NU-DHL-1) on expression profiles accessed via GSE53798. Enrichment plot on SUMOylation signatures, obtained from the Reactome knowledgebase. Adjusted P values (q) are indicated. Volcano plot displays significant gene signatures of the Hallmark gene set (MSigDb) with both MYC Hallmark signatures (V1, V2) highlighted. (E) Enrichment plot on MYC Hallmark signatures, based on the analysis described in (D). Adjusted P values (q) are indicated. (F) Relative viability of Oci-Ly1 cells transduced with a MYC expression plasmid or a control plasmid, treated with increasing SUMOi concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 h (n=3). P values refers to 2,000 nM SUMOi and is determined by unpaired t -test. Immunoblot analysis of Oci-Ly1 control and MYC cell lines.
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Active MYC signaling confers susceptibility to small-molecule SUMO inhibition in diffuse large B-cell lymphoma cell lines. (A) Hierarchical clustering (Euclidean/Ward) of indicated SUMO core components of normalized human diffuse large B-cell lymphoma (DLBCL) transcriptome profiles (GSE98588) revealed SUMO high and SUMO low subgroups as indicated. (B) Gene set enrichment analysis of SUMO high and SUMO low subgroups using Hallmark gene sets from the molecular-signature database (MSigDb). Indicated MYC target gene signatures have been identified using GeneTrail v3 (Kolgomorov-Smirnov-Test). Adjusted P values (q) are indicated. (C) Flow cytometry analysis of relative viability of indicated DLBCL cell lines treated with increasing SUMO inhibitor <t>(SUMOi)</t> <t>concentrations</t> <t>(Tak-981,</t> 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 hours (h) (n=3). (D) Gene set enrichment analysis of SUMOi-responder (Oci-Ly19, SU-DHL-5, SU-DHL-8) vs . SUMOi-non-responder (U-2932, SUDHL-4, DB, NU-DHL-1) on expression profiles accessed via GSE53798. Enrichment plot on SUMOylation signatures, obtained from the Reactome knowledgebase. Adjusted P values (q) are indicated. Volcano plot displays significant gene signatures of the Hallmark gene set (MSigDb) with both MYC Hallmark signatures (V1, V2) highlighted. (E) Enrichment plot on MYC Hallmark signatures, based on the analysis described in (D). Adjusted P values (q) are indicated. (F) Relative viability of Oci-Ly1 cells transduced with a MYC expression plasmid or a control plasmid, treated with increasing SUMOi concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 h (n=3). P values refers to 2,000 nM SUMOi and is determined by unpaired t -test. Immunoblot analysis of Oci-Ly1 control and MYC cell lines.
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Active MYC signaling confers susceptibility to small-molecule SUMO inhibition in diffuse large B-cell lymphoma cell lines. (A) Hierarchical clustering (Euclidean/Ward) of indicated SUMO core components of normalized human diffuse large B-cell lymphoma (DLBCL) transcriptome profiles (GSE98588) revealed SUMO high and SUMO low subgroups as indicated. (B) Gene set enrichment analysis of SUMO high and SUMO low subgroups using Hallmark gene sets from the molecular-signature database (MSigDb). Indicated MYC target gene signatures have been identified using GeneTrail v3 (Kolgomorov-Smirnov-Test). Adjusted P values (q) are indicated. (C) Flow cytometry analysis of relative viability of indicated DLBCL cell lines treated with increasing SUMO inhibitor (SUMOi) concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 hours (h) (n=3). (D) Gene set enrichment analysis of SUMOi-responder (Oci-Ly19, SU-DHL-5, SU-DHL-8) vs . SUMOi-non-responder (U-2932, SUDHL-4, DB, NU-DHL-1) on expression profiles accessed via GSE53798. Enrichment plot on SUMOylation signatures, obtained from the Reactome knowledgebase. Adjusted P values (q) are indicated. Volcano plot displays significant gene signatures of the Hallmark gene set (MSigDb) with both MYC Hallmark signatures (V1, V2) highlighted. (E) Enrichment plot on MYC Hallmark signatures, based on the analysis described in (D). Adjusted P values (q) are indicated. (F) Relative viability of Oci-Ly1 cells transduced with a MYC expression plasmid or a control plasmid, treated with increasing SUMOi concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 h (n=3). P values refers to 2,000 nM SUMOi and is determined by unpaired t -test. Immunoblot analysis of Oci-Ly1 control and MYC cell lines.

Journal: Haematologica

Article Title: Small-molecule SUMO inhibition for biomarker-informed B-cell lymphoma therapy

doi: 10.3324/haematol.2022.280995

Figure Lengend Snippet: Active MYC signaling confers susceptibility to small-molecule SUMO inhibition in diffuse large B-cell lymphoma cell lines. (A) Hierarchical clustering (Euclidean/Ward) of indicated SUMO core components of normalized human diffuse large B-cell lymphoma (DLBCL) transcriptome profiles (GSE98588) revealed SUMO high and SUMO low subgroups as indicated. (B) Gene set enrichment analysis of SUMO high and SUMO low subgroups using Hallmark gene sets from the molecular-signature database (MSigDb). Indicated MYC target gene signatures have been identified using GeneTrail v3 (Kolgomorov-Smirnov-Test). Adjusted P values (q) are indicated. (C) Flow cytometry analysis of relative viability of indicated DLBCL cell lines treated with increasing SUMO inhibitor (SUMOi) concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 hours (h) (n=3). (D) Gene set enrichment analysis of SUMOi-responder (Oci-Ly19, SU-DHL-5, SU-DHL-8) vs . SUMOi-non-responder (U-2932, SUDHL-4, DB, NU-DHL-1) on expression profiles accessed via GSE53798. Enrichment plot on SUMOylation signatures, obtained from the Reactome knowledgebase. Adjusted P values (q) are indicated. Volcano plot displays significant gene signatures of the Hallmark gene set (MSigDb) with both MYC Hallmark signatures (V1, V2) highlighted. (E) Enrichment plot on MYC Hallmark signatures, based on the analysis described in (D). Adjusted P values (q) are indicated. (F) Relative viability of Oci-Ly1 cells transduced with a MYC expression plasmid or a control plasmid, treated with increasing SUMOi concentrations (Tak-981, 0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, 2,000 nM) for 72 h (n=3). P values refers to 2,000 nM SUMOi and is determined by unpaired t -test. Immunoblot analysis of Oci-Ly1 control and MYC cell lines.

Article Snippet: SUMOi (ML-093 and Tak-981, as specified in the figure legends) was either purchased from MedChemExpress or provided by Millennium Pharmaceuticals, Inc., a wholly owned subsidiary of Takeda Pharmaceutical Company Limited.

Techniques: Inhibition, Flow Cytometry, Expressing, Transduction, Plasmid Preparation, Control, Western Blot