Journal: Blood Neoplasia

Article Title: The KDM-family inhibitor JIB-04 sensitizes AML cells to venetoclax by inducing a ferroptosis-like phenotype

doi: 10.1016/j.bneo.2026.100236

Figure Lengend Snippet: JIB-04 inhibits cell proliferation and induces cell death in AML cell lines. (A) OCI-AML3 and MOLM-13 cells were treated with increasing concentrations of the KDM inhibitors CPI-455, GSK-J4, and JIB-04 for 72 hours. Cells were stained with annexin V and PI and analyzed using flow cytometry (mean ± standard deviation [SD]; n = 3). (B) The percentage of viable cells for various AML cell lines after treatment with increasing concentrations of JIB-04 was measured after 72 hours using the MTT assay (mean ± SD; n = 3). IC 50 was calculated using nonlinear regression analysis. (C) Annexin V/PI staining was used to assess the percentage of dead cells after JIB-04 treatment in the indicated concentrations for various AML cell lines (mean ± SD; n = 3). Statistical significance was calculated using ordinary 1-way analysis of variance (ANOVA) with Dunnett multiple-comparison test. (D) Various AML cell lines were stained with anti-CD11b antibody and analyzed by flow cytometry to detect differentiating cells after 72 hours of JIB-04 treatment (mean ± SD; n = 3). Statistical significance was calculated using unpaired t test. ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: AML cells were treated with JIB-04, the antioxidant NAC, the pan caspase inhibitor Z-VAD-FMK (MedChemExpress), or indicated combinations for up to 48 hours.

Techniques: Staining, Flow Cytometry, Standard Deviation, MTT Assay, Comparison

Journal: Blood Neoplasia

Article Title: The KDM-family inhibitor JIB-04 sensitizes AML cells to venetoclax by inducing a ferroptosis-like phenotype

doi: 10.1016/j.bneo.2026.100236

Figure Lengend Snippet: The combination of JIB-04 and VEN is highly synergistic in AML cells. (A) Venetoclax (VEN)-resistant (IC 50 ≥ 1μM) and (B) VEN-sensitive (IC 50 < 1μM) AML cell lines were treated with increasing concentrations of either JIB-04, VEN, or the combination of both (mean ± SD; n = 3). The percentage of dead cells after 72 hours was quantified using annexin V/PI assay. The CI was calculated using the CompuSyn software (CI <1, synergism; CI = 1, additive effects; and CI > 1, antagonism). (C) CD34 + MNC (n = 8) and primary AML samples (n = 5) were treated with 100nM JIB-04, 100nM VEN, or the combination of both for 72 hours. The percentage of apoptotic cells was detected by flow cytometry using the annexin V/PI assay. Statistical significance was calculated using 2-way ANOVA with Tukey multiple-comparison test. (D) OCI-AML3 cells were treated with DMSO, 40nM JIB-04, 1μM VEN, or the combination of both inhibitors for 24 hours and transplanted via tail vein into sublethally irradiated NSG mice. Every group consists of 4 mice. Kaplan-Meier survival plots show survival of mice over time. A statistical analysis was performed using the log-rank (Mantel-Cox) test. ∗ P < .05; ∗∗ P < .01; ∗∗∗∗ P < .0001.

Article Snippet: AML cells were treated with JIB-04, the antioxidant NAC, the pan caspase inhibitor Z-VAD-FMK (MedChemExpress), or indicated combinations for up to 48 hours.

Techniques: Software, Flow Cytometry, Comparison, Irradiation

Journal: Blood Neoplasia

Article Title: The KDM-family inhibitor JIB-04 sensitizes AML cells to venetoclax by inducing a ferroptosis-like phenotype

doi: 10.1016/j.bneo.2026.100236

Figure Lengend Snippet: The KDM inhibitor JIB-04 leads to increased H3K36me3 and upregulation of ferroptosis inducers. (A) Chromatin immunoprecipitation sequencing for H3K36me3 for OCI-AML3 cells treated with 40nM JIB-04 for 24 and 48 hours. Shown are tracks for HMOX1 , SAT1 , and PTGS2 . (B) Volcano plot of significant differentially expressed genes from RNA sequencing analysis of OCI-AML3 cells treated with 40nM JIB-04 at 24 and 48 hours. (C-E) Gene expression analysis of HMOX1 , SAT1 , and PTGS2 by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in OCI-AML3 and MOLM-13 cells treated with 40nM and 50nM JIB-04 for up to 72 hours (mean ± SD; n = 3). β-Actin served as reference gene. mRNA expression was normalized to the DMSO group. Statistical significance was calculated using unpaired t test. (F-G) Protein expression was analyzed by immunoblotting in OCI-AML3 and MOLM-13 cells treated with DMSO or JIB-04 (40nM and 50nM) for 24 hours. β-Actin served as loading control. Data from 3 independent experiments were quantified using the Image Lab software. Protein expression was normalized to the DMSO group. (F) HMOX1, SAT1, and PTGS2 protein expression. (G) E2F8, BRCA1, CDCA2, E2F2, and NRF2 protein expression. (H) Gene ontology (GO) terms overrepresented at 48 hours. (I) GO terms underrepresented at 48 hours. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. tRNA, transfer RNA.

Article Snippet: AML cells were treated with JIB-04, the antioxidant NAC, the pan caspase inhibitor Z-VAD-FMK (MedChemExpress), or indicated combinations for up to 48 hours.

Techniques: ChIP-sequencing, RNA Sequencing, Gene Expression, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Control, Software

Journal: Blood Neoplasia

Article Title: The KDM-family inhibitor JIB-04 sensitizes AML cells to venetoclax by inducing a ferroptosis-like phenotype

doi: 10.1016/j.bneo.2026.100236

Figure Lengend Snippet: JIB-04 treatment causes cell death by ferroptosis in AML cells. (A-B) OCI-AML3 and MOLM-13 cells were treated with 40nM and 50nM JIB-04 for 24 hours (mean ± SD; n = 3). JIB-04 treatment group was normalized to the DMSO group. (A) Mitochondrial ROS were detected using the fluorogenic dye MitoSOX Red and analyzed by flow cytometry. (B) Mitochondrial lipid peroxidation was quantified using the fluorescent probe MitoPerOx and analyzed by flow cytometry. (C-D) OCI-AML3 and MOLM-13 cells were treated with 40nM and 50nM JIB-04 for 48 hours (mean ± SD; n = 3). JIB-04 treatment groups were normalized to the DMSO group. (C) Lipid peroxidation was detected by flow cytometry using the fluorescent probe BODIPY C11. (D) Labile iron was quantified by flow cytometry using the fluorescent probe FerroOrange. In panels A-D, statistical significance was calculated by 2-way ANOVA with Šídák multiple-comparison test. (E-G) Gene expression analysis of HMOX1 , SAT1 , and PTGS2 by qRT-PCR in primary AML cells treated with 100nM JIB-04 for 48 hours (mean ± SD; n = 3). β-Actin served as reference gene. mRNA expression was normalized to the DMSO group. Statistical significance was calculated using unpaired t test. (H-J) Primary AML cells were treated with 100nM JIB-04 for 48 hours and analyzed by flow cytometry for signs of ferroptosis (mean ± SD; n = 3). JIB-04 treatment group was normalized to the DMSO group. (H) MitoSOX red for mitochondrial ROS. (I) MitoPerOx for mitochondrial lipid peroxidation. (J) BODIPY C11 for lipid peroxidation. Statistical significance was calculated by 2-way ANOVA with Šídák multiple-comparison test. ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: AML cells were treated with JIB-04, the antioxidant NAC, the pan caspase inhibitor Z-VAD-FMK (MedChemExpress), or indicated combinations for up to 48 hours.

Techniques: Flow Cytometry, Comparison, Gene Expression, Quantitative RT-PCR, Expressing

Journal: Blood Neoplasia

Article Title: The KDM-family inhibitor JIB-04 sensitizes AML cells to venetoclax by inducing a ferroptosis-like phenotype

doi: 10.1016/j.bneo.2026.100236

Figure Lengend Snippet: Induction of ferroptosis-like phenotype by JIB-04 drives synergism with VEN. (A-C) OCI-AML3 and MOLM-1 cells were treated with 40nM/50nM JIB-04, 8mM/4mM NAC (antioxidant), 25μM Z-VAD-FMK (caspase inhibitor), or the combination JIB-04+NAC and JIB-04+Z-VAD-FMK for up to 48 hours (mean ± SD; n = 3). Cells were stained for ferroptosis markers and analyzed by flow cytometry. JIB-04 treatment group was normalized to the DMSO group. Statistical significance was calculated by 2-way ANOVA with Tukey multiple-comparison test. (A) MitoSOX red for mitochondrial ROS at 24 hours. (B) MitoPerOx for mitochondrial lipid peroxidation at 24 hours. (C) BODIPY C11 for lipid peroxidation at 48 hours. (D) HMOX1 protein levels were visualized by immunoblotting in AML cell lines treated with DMSO, JIB-04, or JIB-04+NAC for 24 hours. (E-F) Quantification of apoptosis by annexin V/PI staining in OCI-AML3 and MOLM-13 cells after treatment with 40nM/50nM JIB-04, 1μM/1nM VEN, 8mM/4mM NAC, or the indicated drug combinations. Statistical significance was calculated by ordinary 1-way ANOVA with Tukey multiple-comparison test. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. Z-VAD, Z-VAD-FMK.

Article Snippet: AML cells were treated with JIB-04, the antioxidant NAC, the pan caspase inhibitor Z-VAD-FMK (MedChemExpress), or indicated combinations for up to 48 hours.

Techniques: Staining, Flow Cytometry, Comparison, Western Blot