tamibarotene  (TargetMol)

Journal: NPJ Breast Cancer

Article Title: Effects of RARα ligand binding domain mutations on breast fibroepithelial tumor function and signaling

doi: 10.1038/s41523-024-00716-5

Figure Lengend Snippet: Mammalian two- hybrid assay of interactions of the wild type or mutant RARα LBD with a fragment of coactivator SRC2, b MED1, a component of the transcription pre-initiation complex and c fragment of corepressor NCOR, treated with increasing concentrations of RA. Left panel: Partially activatable mutants, Right panel: Non-activatable mutants. N = 3, Error bar = SD. d Binding affinities of WT or mutant RARα LBD with fragment of coactivator SRC1 (SRC1-NR2) in the presence of RA and Am80. N = 3, error bar = SE. **p < 0.01.

Article Snippet: Compounds tested include all-trans retinoic acid (known as RA throughout the manuscript, Sigma R2625), Am80 (aka Tamibarotene, Tocris 3507), AM580 (Tocris 0760) and Ch55 (Tocris 2020).

Techniques: Two Hybrid Assay, Mutagenesis, Binding Assay

Journal: NPJ Breast Cancer

Article Title: Effects of RARα ligand binding domain mutations on breast fibroepithelial tumor function and signaling

doi: 10.1038/s41523-024-00716-5

Figure Lengend Snippet: a Overexpression of Flag-tagged RARα WT and mutants in PT024 cells. b Cell viabilities of PT024 WT or mutant RARα expressing cells after treatment with increasing concentration of RA or Am80. N = 3, error bar = SD. *** p < 0.001, **** p < 0.0001. c Cell cycle distribution of PT024 WT or mutant RARα expressing cells when treated with either vehicle or 100 nM Am80. N = 2, error bar = SD. * p < 0.05, **** p < 0.0001. d GSEA analysis shows downregulation of retinoic acid signaling gene targets in mutant RARα-expressing cell lines. e Heatmap of genes associated with reactome retinoic acid signaling pathway. f Quantitative PCR of retinoic acid signaling associated genes. N = 3, error bar = SD. * p < 0.05 ** p < 0.01 *** p < 0.001 **** p < 0.0001.

Article Snippet: Compounds tested include all-trans retinoic acid (known as RA throughout the manuscript, Sigma R2625), Am80 (aka Tamibarotene, Tocris 3507), AM580 (Tocris 0760) and Ch55 (Tocris 2020).

Techniques: Over Expression, Mutagenesis, Expressing, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Determination of differentiation and apoptosis induction following single or combination treatment with revumenib and tamibarotene in different AML cell lines. ( A – D ) Flow cytometry analysis of differentiation ( A , B ) and apoptosis ( C , D ) in MV4:11, MOLM13, OCI-AML3, and HL-60 cells after 72 h incubation with DMSO (0.02% v / v ), 50 nM revumenib, 100 nM tamibarotene, and their combination. The anti-CD11b ratio is calculated relative to the respective isotype control. Graphs represent the mean ± SD from five independent experiments with technical duplicates. Statistical significance was assessed using a two-sided ANOVA (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Flow Cytometry, Incubation, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Time-dependent induction of differentiation and apoptosis following single or combination treatment with revumenib and tamibarotene in different AML cell lines ( A – D ). Flow cytometry analysis of differentiation (upper panel) and apoptosis (lower panel) following 24 h, 48 h, 72 h, and 96 h incubation of MV4:11, MOLM13, OCI-AML3, and HL-60 cells with 0.2% v / v DMSO, 50 nM revumenib, 100 nM tamibarotene, and their combination. CD11b expression for MV4:11 is displayed up to 72 h due to elevated apoptosis rates at 96 h. The anti-CD11b ratio is normalized to the respective isotype control. Graphs depict the mean ± SD from three independent experiments with technical duplicates. Statistical significance was determined using a two-sided ANOVA test (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Flow Cytometry, Incubation, Expressing, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Impact of tamibarotene and revumenib on cellular morphology and surface expression of CD14, CD38, and CD135 ( A , B ). MOLM13 and OCI-AML3 cells were treated with specified concentrations of revumenib, tamibarotene, their combination, and 0.2% v / v DMSO for 4 days. Subsequently, cells were cytospun onto glass slides and stained with hematoxylin and eosin. Representative images were captured with a 40X objective and a CCD camera. ( C , D ) Flow cytometry analysis of CD14 expression after 24 h and 72 h incubation of MV4:11, MOLM13, and OCI-AML3 cells with specified concentrations of revumenib, tamibarotene, their combination, and 0.2% v / v DMSO, normalized to the respective isotype control. ( E , F ) Flow cytometry analysis of anti-CD135 and anti-CD38 staining after 24 h incubation for indicated conditions. Antibody ratios are normalized to the respective isotype control. Graphs represent the mean ± SD from three independent experiments with technical duplicates. Statistical significance was determined using a two-sided ANOVA test (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Expressing, Staining, Flow Cytometry, Incubation, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Western blot analysis of differentiation and apoptosis proteins in response to revumenib and tamibarotene treatments. Protein lysates from MV4:11 ( A ), MOLM13 ( B , C ), OCI-AML3 ( D ), and HL-60 ( E ) cells—treated for 24 h or 72 h with DMSO 0.2% v / v , 50 nM revumenib, 100 nM tamibarotene and their combination—underwent immunoblotting to assess the expression and phosphorylation status of specified proteins. GAPDH levels were detected as a loading control. Corresponding densitometry analysis was conducted, and fold changes normalized to the loading control and DMSO control are presented within the blots. Representative blots from three independent experiments are displayed. Next to the blots, the quantification of densitometry analysis is summarized from all replicates (at least two representative blots per protein) and indicated as log 2 fold change. The uncropped blots are shown in . Except for ( F ), where the p-CEBPA-to-CEBPA ratio is illustrated, all proteins are normalized to the DMSO control, which is set to 1 and therefore not shown. Statistical significance was determined using a two-sided ANOVA (*** p < 0.001; ** p < 0.01; * p < 0.05; ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Western Blot, Expressing, Phospho-proteomics, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Synergistic reduction in metabolic activity with the combination of revumenib and tamibarotene ( A – D ). Indicated cell lines were incubated for 48 h with concentration series of revumenib (0, 10, 50, 100 nM) and tamibarotene (0, 50, 100, 300, 500 nM). Metabolic activity was assessed by PrestoBlue viability assay in a 96-well plate, and results were normalized to the DMSO control. Graphs represent the mean ± SD from two independent experiments in technical triplicates. Statistical significance was determined using a two-sided ANOVA (ns, not significant). ( E – H ) Cell lines were incubated for 48 h with a revumenib concentration series (0, 50, 100, 300, and 500 nM), either in combination with DMSO (0.01% v / v ) or 50 nM tamibarotene. Metabolic activity was measured by PrestoBlue viability assay and graphs depict the mean ± SD from three independent experiments in technical triplicates. Data were analyzed using SynergyFinder software (Version 3) to calculate ZIP synergy scores (0–10 additive effects, >10 synergistic effects). Corresponding heat maps are illustrated on the right side.

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Activity Assay, Incubation, Concentration Assay, Viability Assay, Control, Software

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Effects of revumenib and tamibarotene on primary AML cells. PBMCs were thawed and rested in IMDM medium for 24 h before incubating with 50 nM revumenib, 100 nM tamibarotene, and DMSO (0.02% v / v ) for 72 h. Subsequently, protein lysates underwent immunoblotting for specified proteins, quantified by densitometry, and normalized to the loading and DMSO control. Flow cytometry measured Annexin V, CD11b, CD34, and CD38 surface expression (all presented as ratios to isotype control from geometric means) after 72 h of incubation. The uncropped blots are shown in .

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Western Blot, Control, Flow Cytometry, Expressing, Incubation

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Determination of differentiation and apoptosis induction following single or combination treatment with revumenib and tamibarotene in different AML cell lines. ( A – D ) Flow cytometry analysis of differentiation ( A , B ) and apoptosis ( C , D ) in MV4:11, MOLM13, OCI-AML3, and HL-60 cells after 72 h incubation with DMSO (0.02% v / v ), 50 nM revumenib, 100 nM tamibarotene, and their combination. The anti-CD11b ratio is calculated relative to the respective isotype control. Graphs represent the mean ± SD from five independent experiments with technical duplicates. Statistical significance was assessed using a two-sided ANOVA (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Flow Cytometry, Incubation, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Time-dependent induction of differentiation and apoptosis following single or combination treatment with revumenib and tamibarotene in different AML cell lines ( A – D ). Flow cytometry analysis of differentiation (upper panel) and apoptosis (lower panel) following 24 h, 48 h, 72 h, and 96 h incubation of MV4:11, MOLM13, OCI-AML3, and HL-60 cells with 0.2% v / v DMSO, 50 nM revumenib, 100 nM tamibarotene, and their combination. CD11b expression for MV4:11 is displayed up to 72 h due to elevated apoptosis rates at 96 h. The anti-CD11b ratio is normalized to the respective isotype control. Graphs depict the mean ± SD from three independent experiments with technical duplicates. Statistical significance was determined using a two-sided ANOVA test (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Flow Cytometry, Incubation, Expressing, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Impact of tamibarotene and revumenib on cellular morphology and surface expression of CD14, CD38, and CD135 ( A , B ). MOLM13 and OCI-AML3 cells were treated with specified concentrations of revumenib, tamibarotene, their combination, and 0.2% v / v DMSO for 4 days. Subsequently, cells were cytospun onto glass slides and stained with hematoxylin and eosin. Representative images were captured with a 40X objective and a CCD camera. ( C , D ) Flow cytometry analysis of CD14 expression after 24 h and 72 h incubation of MV4:11, MOLM13, and OCI-AML3 cells with specified concentrations of revumenib, tamibarotene, their combination, and 0.2% v / v DMSO, normalized to the respective isotype control. ( E , F ) Flow cytometry analysis of anti-CD135 and anti-CD38 staining after 24 h incubation for indicated conditions. Antibody ratios are normalized to the respective isotype control. Graphs represent the mean ± SD from three independent experiments with technical duplicates. Statistical significance was determined using a two-sided ANOVA test (ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Expressing, Staining, Flow Cytometry, Incubation, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Western blot analysis of differentiation and apoptosis proteins in response to revumenib and tamibarotene treatments. Protein lysates from MV4:11 ( A ), MOLM13 ( B , C ), OCI-AML3 ( D ), and HL-60 ( E ) cells—treated for 24 h or 72 h with DMSO 0.2% v / v , 50 nM revumenib, 100 nM tamibarotene and their combination—underwent immunoblotting to assess the expression and phosphorylation status of specified proteins. GAPDH levels were detected as a loading control. Corresponding densitometry analysis was conducted, and fold changes normalized to the loading control and DMSO control are presented within the blots. Representative blots from three independent experiments are displayed. Next to the blots, the quantification of densitometry analysis is summarized from all replicates (at least two representative blots per protein) and indicated as log 2 fold change. The uncropped blots are shown in . Except for ( F ), where the p-CEBPA-to-CEBPA ratio is illustrated, all proteins are normalized to the DMSO control, which is set to 1 and therefore not shown. Statistical significance was determined using a two-sided ANOVA (*** p < 0.001; ** p < 0.01; * p < 0.05; ns, not significant).

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Western Blot, Expressing, Phospho-proteomics, Control

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Synergistic reduction in metabolic activity with the combination of revumenib and tamibarotene ( A – D ). Indicated cell lines were incubated for 48 h with concentration series of revumenib (0, 10, 50, 100 nM) and tamibarotene (0, 50, 100, 300, 500 nM). Metabolic activity was assessed by PrestoBlue viability assay in a 96-well plate, and results were normalized to the DMSO control. Graphs represent the mean ± SD from two independent experiments in technical triplicates. Statistical significance was determined using a two-sided ANOVA (ns, not significant). ( E – H ) Cell lines were incubated for 48 h with a revumenib concentration series (0, 50, 100, 300, and 500 nM), either in combination with DMSO (0.01% v / v ) or 50 nM tamibarotene. Metabolic activity was measured by PrestoBlue viability assay and graphs depict the mean ± SD from three independent experiments in technical triplicates. Data were analyzed using SynergyFinder software (Version 3) to calculate ZIP synergy scores (0–10 additive effects, >10 synergistic effects). Corresponding heat maps are illustrated on the right side.

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Activity Assay, Incubation, Concentration Assay, Viability Assay, Control, Software

Journal: Cancers

Article Title: Synergistic Effects of the RAR alpha Agonist Tamibarotene and the Menin Inhibitor Revumenib in Acute Myeloid Leukemia Cells with KMT2A Rearrangement or NPM1 Mutation

doi: 10.3390/cancers16071311

Figure Lengend Snippet: Effects of revumenib and tamibarotene on primary AML cells. PBMCs were thawed and rested in IMDM medium for 24 h before incubating with 50 nM revumenib, 100 nM tamibarotene, and DMSO (0.02% v / v ) for 72 h. Subsequently, protein lysates underwent immunoblotting for specified proteins, quantified by densitometry, and normalized to the loading and DMSO control. Flow cytometry measured Annexin V, CD11b, CD34, and CD38 surface expression (all presented as ratios to isotype control from geometric means) after 72 h of incubation. The uncropped blots are shown in .

Article Snippet: Tamibarotene and revumenib were purchased from Selleck Chemicals and dissolved in DMSO.

Techniques: Western Blot, Control, Flow Cytometry, Expressing, Incubation