palbociclib  (MedChemExpress)

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Experimental design for the drug screening: MV4-11 cells cocultured with HS-5 stromal cells were pretreated for 24 h with either vehicle control or palbociclib, followed by the exposure to either the second molecule alone, or the combination of palbociclib and the second molecule for an additional 72 h. Cells were then harvested to assess the expression of the myeloid differentiation marker CD11b by flow cytometry and to quantify the minor compartment of colony-forming cells. ( B ) List of molecules used in the screen (first column), and results of the analyses of CD11b expression following monotherapies (second column) or combined treatments with palbociclib (third column). Heatmap was applied to highlight the increase in CD11b expression. ( C ) Results of the colony-forming cell assay for monotherapies involving TCP and ATRA or in combination with palbociclib. Colonies (CFU-L) are expressed as numbers (second column) and as the percentage of colonies relative the control cells (last column). Heatmap was used in the table to highlight the drop in colony number.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Drug discovery, Control, Expressing, Marker, Flow Cytometry

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Experimental design for the treatment of AML cells. AML cells, cocultured with HS-5 stromal cells, were treated for 4 days with palbociclib at 0.5 μM and/or TCP at 5 μM, and analyzed by flow cytometry and May-Grünwald–Giemsa staining. ( B ) The percentage of MV4-11 cells expressing CD11b ( n = 3), CD14 ( n = 3) or CD86 ( n = 4) myeloid markers was quantified by flow cytometry following the indicated treatments. Ctrl control, TCP tranylcypromine, Palbo palbociclib, Combo is the combination of palbociclib with tranylcypromine. The exact adjusted p values were as follows: Left panel, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0001. Middle panel: Ctrl vs Combo p = 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0008. Right panel: Ctrl vs Combo p = 0.0002, TCP vs Combo p = 0.0003, Palbo vs Combo p = 0.0023. ( C – E ) Expression of the late monocytic marker CD86 on MOLM-14 ( C ) ( n = 3), PL-21 ( D ) ( n = 3), and THP-1 ( E ) ( n = 3) AML cell lines. The percentage of cells expressing CD86 was quantified by flow cytometry following the indicated treatments. The exact adjusted p values were as follows: ( C ) Ctrl vs Combo p = 0.0003, TCP vs Combo p = 0.0005, Palbo vs Combo p = 0.0019. ( D ) Ctrl vs Combo p < 0.0001, TCP vs Combo p < 0.0001, Palbo vs Combo p = 0.0117, Ctrl vs Palbo p = 0.0005, TCP vs Palbo p = 0.0012. ( E ) Ctrl vs Combo p = 0.0005, TCP vs Combo p = 0.0019, Palbo vs Combo p = 0.0098, Ctrl vs Palbo p = 0.0297. ( F ) Cell morphology of the AML cell lines following the treatments. AML cell lines were stained with May-Grünwald–Giemsa. Cells were observed under a Zeiss Apotome microscope. The indicated scale is 50 µm. ( G ) Primary AML patient cells were treated with the indicated treatments for 96 h. After treatment, cells were analyzed by flow cytometry for the expression of CD11b ( n = 17) and CD86 ( n = 14). Each colored dot represents a sample. Exact adjusted pvalues: left panel, Ctrl vs TCP p = 0.0070, Ctrl vs Palbo p = 0.0059, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0004, Palbo vs Combo p = 0.0005; right panel, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0022. ( H ) May-Grünwald–Giemsa staining of five primary AML patient samples following the indicated treatments. The indicated scale is 50 µm. Histograms indicate the mean of independent experiments ± SD. Statistical analyses were performed using a one-way ANOVA and followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Flow Cytometry, Staining, Expressing, Control, Marker, Microscopy

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Top panel: Experimental design to quantify colony-forming cells of treated AML cells. MV4-11 cells were treated for 4 days with palbociclib (0.5 μM), TCP (5 μM), or the combination of palbociclib and TCP. Bottom panel: Representative images of methylcellulose colony formation assays and the quantification of the colonies (mean ± SD; n = 6), relative to control. Exact adjusted p values: Ctrl vs TCP p < 0.0001; Ctrl vs Palbo p < 0.0001; Ctrl vs Combo p < 0.0001; TCP vs Combo p < 0.0001; Palbo vs Combo p < 0.0001. ( B , C ) Quantification of the colonies on MOLM-14 and PL-21 AML cells, treated as described in ( A ), relative to control. Data represent the mean ± SD of n = 3 independent experiments. In ( B ), adjusted p values: Ctrl vs Combo p = 0.0044, TCP vs Combo p = 0.0030, Palbo vs Combo p = 0.0056. In ( C ), Ctrl vs Palbo p = 0.0005, Ctrl vs Combo p = 0.0001, TCP vs Palbo p = 0.0052, TCP vs Combo p = 0.0006. ( D , E ) Effects of the indicated treatments on CFCs of primary patient AML cells. ( D ) Adult samples ( n = 24); ( E ) pediatric samples ( n = 8). Cells treated with the indicated molecules for 96 h were then seeded in methylcellulose and colonies counted after 10 days. The number of colonies is represented as a relative number of colonies compared to control cells. Scatter plots indicate the median. Each colored dot represents a sample. The exact adjusted pvalues were: ( D ) Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001, TCP vs Combo p < 0.0001, Palbo vs Combo p < 0.0001 ; ( E ) Ctrl vs TCP p < 0.0001, Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001, TCP vs Palbo p = 0.0035, TCP vs Combo p < 0.0001, Palbo vs Combo p = 0.0002. ( F ) Secondary replating assays from colonies obtained in ( D ) for six adult patient samples ( n = 6). Scatter plots indicate the median, with each colored dot representing a sample. The exact adjusted p values: Ctrl vs Combo p = 0.0009; TCP vs Palbo p = 0.0078; Palbo vs Combo p = 0.0001. Statistical analyses were performed using a one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Control

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A , B ) Alternative LSD1 or CDK6 inhibitors. MV4-11 cells were treated with the LSD1 inhibitor ORY-1001 (ORY) with palbociclib ( A ) or with TCP and the CDK6 inhibitor ribociclib (Ribo) ( B ). After the 4-day treatment, cells were stained with May-Grünwald–Giemsa (top panel), and their capacity to form colonies was evaluated (bottom panel). ( C ) MV4-11 cells cocultured with HS-5 stromal cells were incubated with CDK6 inhibitor ribociclib at 1 μM, with ORY-1001 at 0.5 nM, or with the combination of ribociclib and ORY-1001 (Combo) for 96 h. After the treatment, leukemic cells were seeded in equal numbers in methylcellulose for 10 days. Data represent the mean ± SD of n = 3 independent experiments. Statistical analyses were performed using a one-way ANOVA followed by Tukey’s test. ** p < 0.01, *** p < 0.001, **** p < 0.0001. Adjusted pvalues: Ctrl vs ORY p = 0.0075, Ctrl vs Ribo p < 0.0001, Ctrl vs Combo p < 0.0001, ORY vs Ribo p = 0.0018, ORY vs Combo p = 0.0003. ( D ) Evaluation of the synergy of CDK6 and LSD1 inhibitor combinations. MV4-11 cells were treated with increased concentrations of CDK6 inhibitor (palbociclib 0.125 to 1 μM; ribociclib 0.25 to 2 μM; abemaciclib 6.25 to 50 nM) in combination with LSD1 inhibitor (TCP 1.25 to 10 μM; ORY-1001 0.125 to 1 nM; GSK2879552 1.25 to 10 μM) for 96 h. The percentage of cells expressing CD11b was quantified by flow cytometry. Results were analyzed using the SynergyFinder tool, and represented by 2D contour plots. The red color indicates synergy.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Staining, Incubation, Expressing, Flow Cytometry

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) MV4-11 cells transfected with control (Ctrl) or LSD1 siRNAs were incubated with palbociclib (0.5 μM), as indicated, for 4 days. The percentage of CD11b-expressing cells was assessed by flow cytometry (left panel). Data represent the mean ± SD of n = 3 independent experiments. LSD1 protein expression was quantified by Western blot analysis (right panel); one of three experiments is shown. Statistical analyses were performed using a one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001. Adjusted pvalues: siCtrl vs siLSD1 p = 0.0363, siCtrl vs siLSD1+Palbo p = 0.0006, siLSD1 vs siLSD1+Palbo p = 0.0131, siCtrl+Palbo vs siLSD1+Palbo p = 0.0020. ( B ) MV4-11 cells transfected with control (Ctrl) or CDK6 siRNAs were incubated with 5 μM TCP or 0.5 nM ORY-1001, as indicated, for 4 days. The percentage of CD11b-expressing cells was assessed by flow cytometry (left panel). Data represent the mean ± SD of n = 2 independent experiments. CDK6 protein expression was quantified by Western blot. One representative experiment out of two is shown (right panel). ( C ) MV4-11 cells transfected with control, CDK6 and/or LSD1 siRNAs were seeded on methylcellulose. CFCs were analyzed at day 10 post plating.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

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

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Bioluminescent imaging of NSG mice at day 24, post-xenotransplantation of 2 × 10 5 luciferase-labeled MOLM-14 cells. Mice were treated by intraperitoneal injections of either palbociclib (25 mg/kg), ORY-1001 (0.0125 mg/kg), the combination of the two molecules or vehicle control, for 4 weeks. ( B , C ) Quantification of the percentage of MOLM-14 cells defined as hCD45 + hCD33+ cells, by flow cytometry, in the bone marrow ( B ) and in the spleen ( C ). Data represent the mean ± SD of one experiment with n = 5 mice per condition. The adjusted pvalues were: ( B ) Ctrl vs ORY p = 0.0057, Ctrl vs Combo p < 0.0001, ORY vs Palbo p = 0.0018, Palbo vs Combo p < 0.0001 ; ( C ) Ctrl vs ORY p = 0.0055, Ctrl vs Combo p = 0.0012, Palbo vs Combo p = 0.0337. ( D ) Quantification of hCD45 + hCD33+ leukemic blast cells in NSG mice transplanted with primary AML patient cells. Treatment with the combination of palbociclib and ORY-1001 was initiated once the engraftment was confirmed by the appearance of blast cells in the bloodstream, and were administered for 3–6 weeks until an endpoint was reached. The percentage of human myeloid cells was assessed by flow cytometry in the bone marrow. Data represent the mean ± SD. Statistical analysis was performed using an unpaired one-tailed Student’s t -test. PDX#1 p < 0.0001 ; PDX#2 p < 0.0001 ; PDX#3 p = 0.3301 ; PDX#4 p = 0.0033 ; PDX#5 p = 0.0368. ( E , F ) Flow cytometry quantification of hCD45 + hCD33+ leukemic blast cells in blood, bone marrow and spleen of NSG mice engrafted with PDX#2 ( E ) and PDX#4 ( F ), treated as indicated. Scatter plots indicate the mean ± SD of an experiment with n = 10 mice per condition. Adjusted p values in ( E ): Left panel, Ctrl vs ORY p < 0.0001, Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001 ; Middle panel, Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001, ORY vs Palbo p < 0.0001, ORY vs Combo p < 0.0001, Palbo vs Combo p = 0.0004; Right panel, Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001, ORY vs Palbo p < 0.0001, ORY vs Combo p < 0.0001, Palbo vs Combo p = 0.0030. ( F ) Left panel, Ctrl vs ORY p = 0.0033, Ctrl vs Palbo p = 0.0230, Ctrl vs Combo p < 0.0001; Middle panel, Ctrl vs Combo p = 0.0047; Right panel, Ctrl vs ORY p = 0.0199, Ctrl vs Palbo p < 0.0001, Ctrl vs Combo p < 0.0001, ORY vs Palbo p = 0.0449, ORY vs Combo p = 0.0049. Except for ( D ), statistical analyses were performed using a one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns for not significant.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Imaging, Luciferase, Labeling, Control, Flow Cytometry, One-tailed Test

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Evaluation of the combined treatment on mouse hematopoiesis. C57BL/6 mice were treated with intraperitoneal injections of the vehicle control or a combination of palbociclib (25 mg/kg) and ORY-1001 (0.0125 mg/kg), 4 days a week for 4 weeks. At the end of the treatments, blood counts and the bone marrow myeloid progenitors were analyzed. Data represent the mean ± SD of n = 5 mice. Statistical analysis was performed using an unpaired two-tailed Student’s t -test. ns is not significant. * p < 0.05. CMP common myeloid progenitors, GMP granulocyte-monocyte progenitors, MEP megakaryocyte-erythroid progenitors. Top panels, p = 0.0163, p = 0.0254, p = 0.0519, and p = 0.0211. Bottom panels, p = 0.6117, p = 0.4861, p = 0.8805, p = 0.7955, p = 0.0995, and p = 0.1129. ( B ) Quantification of hCD45 + hCD33+ leukemic blast cells in the spleen and blood of the PDX NSG mouse models, as shown in Fig. . PDX#1 n = 5, PDX#2 n = 5, PDX#3 n = 5, PDX#4 n = 5, PDX#5 n = 3. Statistical analysis was performed using an unpaired one-tailed Student’s t -test. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( C ) Analysis of the leukemic cells remaining following in vivo treatment of PDX mouse models. (Left) The expression of CD86 and CD163 late myeloid markers were analyzed on the remaining leukemic cells (detected as hCD45+ cells), for each untreated and matched PDX treated pair. (Middle and Right) Evaluation of the number of CFC progenitors remaining in the bone marrow of four independent PDX samples. Cells were seeded in methylcellulose in equal numbers for 10 days. The right histogram combined results from the four PDX, expressed as a percentage of colonies obtained relative to the control cells. Data represent the mean ± SD of n = 4 samples per condition. Statistical analysis was performed using a paired one-tailed Student’s t -test, **** p < 0.0001.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: Control, Two Tailed Test, One-tailed Test, In Vivo, Expressing

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: RNA-seq analyses of MV4-11 cells treated for 4 days with the combination or the single molecules as indicated. ( A ) Volcano plots highlighting the differentially expressed genes (DEG) following treatments ( n = 2) with either TCP, palbociclib or the combination, compared to control cells ( p adj < 0.01). Statistical significance was assessed using the Wald test, and the resulting p values were adjusted for multiple testing using the Benjamini–Hochberg procedure to control the false discovery rate. ( B ) Venn diagram indicating the number of DEG compared to the control (log2 fold change > 1, p adj < 0.01). ( C ) Gene set enrichment analyses (GSEA) of myeloid differentiation and LSC signature, comparing the combination treatment to the control. Statistical analyses were performed using the GSEA software. ( D ) GSEA of the LSD1 inhibition signature, comparing the cells treated with the combination with TCP only. Statistical analyses were performed using the GSEA software. ( E ) Biological processes associated with the deregulated genes, using GeneAnalytics.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: RNA Sequencing, Control, Software, Inhibition

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: RNA-seq analysis of 6 primary AML human samples treated either with the combination (0.5 μM palbociclib and 5 μM TCP) or the single molecules (TCP or Palbo) for 96 h. ( A ) Volcano plots illustrating the significantly deregulated genes for each treatment compared to the control cells (log2 fold change > 1, p adj < 0.01) of primary patient samples ( n = 6). Statistical significance was assessed using Wald test, and the resulting p values were adjusted for multiple testing using the Benjamini–Hochberg procedure to control the false discovery rate. ( B ) Venn diagrams indicating the number of significantly deregulated genes (left panel), and the number of down- and up-regulated genes compared to the control cells (middle and right panels). The thresholds were log2 fold change > 1, and p adj < 0.01. ( C , D ) GSEA of myeloid differentiation ( C ) and LSC/immature signatures ( D ) comparing the combination treatment to the control. ( E ) GSEA of the LSD1 signature, comparing the combination treatment to TCP. ( F , G ) Heatmaps of the differentiation signature ( F ) and the LSD1 signature ( G ) for each individual patient sample. Normalized TPM values are represented as Z-scores.

Article Snippet: Palbociclib monohydrochloride (in vivo studies) , MedChemExpress , HY-50767A.

Techniques: RNA Sequencing, Control