Figure 5.
Menin inhibitor resistance in PRC1.1-deficient AML cells is mediated by aberrant reactivation of type II menin target MYC. (A-C) Volcano plots showing GSEA results using the Molecular Signatures Database Transcription Factor Targets gene sets, comparing VTP50469-treated OCI-AML2 cells with sgPCGF1 (A, left) or sgBCOR (A, right) with sgLuc controls, and similarly for MV4-11 (B) and MOLM-13 (C) cells with sgPCGF1, all under VTP50469 treatment. (D) Bar plots showing the RNA-seq normalized counts of the MYC gene in indicated AML cells. Adjusted P values were calculated using DESeq2. (E) GSEA showing depletion of MYC gene signature in OCI-AML2, MV4-11, MOLM-13, and PDX MLL-AF9 cells (DFAM68555 characterized as MLL-AF9 fusion, MLL3 wild-type AML, and further treated with vehicle [DMSO] or a menin inhibitor [VTP50469]) 11 after menin inhibition treatment. (F) Scatterplot representing the positive correlations between MYC and MEN1 expression levels in samples of patient with AML from the Beat AML and The Cancer Genome Atlas (TCGA) studies, along with AML cell lines from the DepMap database. (G) Schematic diagram displaying the competitive proliferation assay for assessing the effect of MYC constitutive expression on AML cell response to menin inhibition. (H-J) Bar plots showing the percentages of MYC-RFP+ cells in OCI-AML2 (H; 100 nM), MV4-11 (I; 30 nM), and MOLM-13 (J; 10 nM) AML cell lines treated with either DMSO or VTP50469 over specified time periods. (K-M) Bar plots showing the percentages of CD11b+ cells in OCI-AML2 (K), MV4-11 (L), and MOLM-13 (M) AML cell lines with or without MYC overexpression, under treatment with either DMSO or VTP50469 for 6 days. (N) Dose-response curves showing the viability of OCI-AML2 cells expressing the indicated sgRNAs after a 6-day treatment with DMSO control or various doses of VTP50469. All cell viabilities were normalized to DMSO treatment. Calculated IC50 values were shown in boxplots. (O) Dose-response curves showing viabilities of OCI-AML2 cells expressing either sgLuc control, sgPCGF1 alone, or sgPCGF1 combined with sgMYC, after a 9-day treatment with DMSO control or various doses of VTP50469. All cell viabilities were normalized to DMSO treatment. Calculated IC50 values were shown in boxplots. (P) Bar plots showing cell viability of OCI-AML2 cells expressing either sgLuc or sgPCGF1 under 9-day treatment with DMSO, 100 nM VTP50469, 1 nM ABBV075, or a combination of VTP50469 and ABBV075 (combo). (Q) Bar plots showing cell viability of the indicated mouse RN2c cells under a 4-day treatment with DMSO, 100 nM VTP50469, 1 nM ABBV075, or a combination of VTP50469 and ABBV075 (combo). FDR, false discovery rate; NES, normalized enrichment score; ∗∗∗∗P < .0001.

Menin inhibitor resistance in PRC1.1-deficient AML cells is mediated by aberrant reactivation of type II menin target MYC. (A-C) Volcano plots showing GSEA results using the Molecular Signatures Database Transcription Factor Targets gene sets, comparing VTP50469-treated OCI-AML2 cells with sgPCGF1 (A, left) or sgBCOR (A, right) with sgLuc controls, and similarly for MV4-11 (B) and MOLM-13 (C) cells with sgPCGF1, all under VTP50469 treatment. (D) Bar plots showing the RNA-seq normalized counts of the MYC gene in indicated AML cells. Adjusted P values were calculated using DESeq2. (E) GSEA showing depletion of MYC gene signature in OCI-AML2, MV4-11, MOLM-13, and PDX MLL-AF9 cells (DFAM68555 characterized as MLL-AF9 fusion, MLL3 wild-type AML, and further treated with vehicle [DMSO] or a menin inhibitor [VTP50469]) 11 after menin inhibition treatment. (F) Scatterplot representing the positive correlations between MYC and MEN1 expression levels in samples of patient with AML from the Beat AML and The Cancer Genome Atlas (TCGA) studies, along with AML cell lines from the DepMap database. (G) Schematic diagram displaying the competitive proliferation assay for assessing the effect of MYC constitutive expression on AML cell response to menin inhibition. (H-J) Bar plots showing the percentages of MYC-RFP+ cells in OCI-AML2 (H; 100 nM), MV4-11 (I; 30 nM), and MOLM-13 (J; 10 nM) AML cell lines treated with either DMSO or VTP50469 over specified time periods. (K-M) Bar plots showing the percentages of CD11b+ cells in OCI-AML2 (K), MV4-11 (L), and MOLM-13 (M) AML cell lines with or without MYC overexpression, under treatment with either DMSO or VTP50469 for 6 days. (N) Dose-response curves showing the viability of OCI-AML2 cells expressing the indicated sgRNAs after a 6-day treatment with DMSO control or various doses of VTP50469. All cell viabilities were normalized to DMSO treatment. Calculated IC50 values were shown in boxplots. (O) Dose-response curves showing viabilities of OCI-AML2 cells expressing either sgLuc control, sgPCGF1 alone, or sgPCGF1 combined with sgMYC, after a 9-day treatment with DMSO control or various doses of VTP50469. All cell viabilities were normalized to DMSO treatment. Calculated IC50 values were shown in boxplots. (P) Bar plots showing cell viability of OCI-AML2 cells expressing either sgLuc or sgPCGF1 under 9-day treatment with DMSO, 100 nM VTP50469, 1 nM ABBV075, or a combination of VTP50469 and ABBV075 (combo). (Q) Bar plots showing cell viability of the indicated mouse RN2c cells under a 4-day treatment with DMSO, 100 nM VTP50469, 1 nM ABBV075, or a combination of VTP50469 and ABBV075 (combo). FDR, false discovery rate; NES, normalized enrichment score; ∗∗∗∗P < .0001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal