Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: Relative mRNA levels of major enzymes in the fatty acid synthesis pathway from FLT3-ITD cell lines ( A ) and FLT3-WT cell lines ( B ). Ctrl, control cells without treatment. Quizar_6 h, treatment with quizartinib (10 nM) for 6 h. Quizar_12 h, treatment with quizartinib (10 nM) for 12 h. Bars, means ± SD, n = 3 per group. C KEGG metabolism related pathways of downregulated genes in CD45 positive cells after quizartinib treatment in GSE202222 dataset. Arrows, lipid metabolism pathways. D Gene set enrichment analysis of the KEGG signature fatty acid metabolism (KEGG ID: hsa01212) after quizartinib treatment in GSE202222 dataset. E Volcano plot of differentially expressed genes in GSE202222. Labeled genes were those downregulated in the fatty acid or lipid metabolism after quizartinib treatment ( p < 0.05, |Fold Change| > 1.5).

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Control, Labeling

Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: Relative Srebf1 / SREBF1 mRNA level in murine ( A ) and human ( B ) leukemia cell lines with control, quizartinib (10 nM) treatment for 6 h and 12 h. N = 3 replicates. Bars, means ± S.D. C Upper panel, western blotting of SREBP1 and FASN in FLT3-WT and FLT3-ITD cell lines before and after quizartinib treatment of 6 and 12 h. P precursor form, N nuclear form. Lower panel, comparison of SREBP1 protein expression in the FLT3/ITD cells before and after quizartinib treatment. N = 3 replicates. Bars, means ± S.D. D Log2CPM gene-expression values of SREBF1 / Srebf1 in TCGA_LAML, GSE163926 and GSE202222 datasets. E The polysome profile of BaF3/ITD cells. 40S, 60S, and 80S ribosomal subunits and polysomes were fractionated using 5–50% density gradient sucrose buffer and monitored with A260 measurements. F , G Total RNA of BaF3/ITD cells before and after quizartinib treatment was extracted from each fraction. The expression of Srebf1 and Actb was measured by qPCR. The mRNA content from monosomes (40S, 60S, 80S) represent the non-translated portion. The mRNA content from polysome represents the translated portion. N = 3 independent experiments. Bars, means ± S.D.

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Control, Western Blot, Comparison, Expressing, Gene Expression

Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: A BaF3/ITD were pre-treated with vehicle control (DMSO) or quizartinib (10 nM) for 6 h. SREBP1 (precursor) protein expression was then detected after treatment with cycloheximide (CHX, 100 μg/mL) at the indicated time points. B The plotted graph of three independent experiments showing the relative amount of SREBP1 at each time point compared to the control without CHX treatment. C Western blotting showing SREBP1 protein expression in the presence of CHX (100 μg/mL) and MG132 (10 μM). D Expression of total AKT, phosphor-AKT (p-AKT S473 ), SREBP1 and FASN in BaF3/ITD and MV4-11 cells before and after treatment of MK2206 (5 μM) for 6 and 12 h. E Expression of total AKT, phospho-AKT (p-AKT S473 ), total GSK3β and phospho-GSK3β (p-GSK3β S9 ) in BaF3/ITD and MV4-11 cells before and after treatment of quizartinib (10 nM) for 6 and 12 h. Vinculin was used as loading control. F Expression of SREBP1 and FASN in BaF3/ITD and MV4-11 cells before and after treatment of CHIR-99021 (3 μM) for 6 and 12 h. G The ubiquitination and phosphorylation (pan phospho-Ser/Thr) levels of SREBP1 before and after treatment of CHIR-99021 for 6 h. IP immunoprecipitation. Total input of SREBP1 and α-Tubulin was shown as equal loading. H Expression of SREBP1in BaF3/ITD and MV4-11 cells with shRNA targeting GSK3β . NC non-specific control. Sh #1 and Sh #2, different sequencing of shRNA targeting GSK3β . I The ubiquitination and phosphorylation (pan phospho-Ser/Thr) levels of SREBP1 before and after treatment of quizartinib for 6 h. J An illustrated pathway displaying the regulation of protein degradation of SREBP by the FLT3/AKT/GSKβ phosphorylation cascade and the kinase inhibitors including quizartinib, MK2206 and CHIR-99021.

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Control, Expressing, Western Blot, Immunoprecipitation, shRNA, Sequencing

Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: A , B The FLT3-ITD leukemia cell lines were treated with vehicle or quizartinib (10 nM) at the indicated time points. Mitochondrial membrane potential and cell death was assessed by Rho123 and Annexin V/PI, respectively. n = 3 per group. Bars, means ± S.D. C Lipidomic analysis of MOLM-13 cells before and after treatment of quizartinib (10 nM) for 12 h. Significant differentially abundant metabolites between two groups ( p < 0.05, |Log2FC | > 1 and VIP > 1) are shown in red (upregulated) and blue (downregulated) in the volcano plot. D The pie graph illustrating the proportion of each lipid class contributing to the total number of upregulated (red) and downregulated (blue) features. E The illustration of enzymatic steps involved in biosynthesis of major phospholipids. F Comparison of lipid abundance between the control and treatment group in all lipid classes with statistical significance. Blue stars, major types of phospholipids involved in the biosynthesis pathway. N = 6 replicates. Bars, means ± S.D. ***( p < 0.001) and ****( p < 0.0001) indicate p values for the major phospholipids with blue stars. p < 0.05 for all other lipid species. G The absolute cardiolipin contents in FLT3/ITD cells treated before and after quizartinib (10 nM, 12 h) were detected by the fluorometric assay as described in the method. N = 3 replicates. Bars, means ± S.D. The abbreviations of all lipid species and enzymes were listed in the Supplementary Table .

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Membrane, Comparison, Control

Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: A , B Quantitative analysis of cell death after different drug treatment for 48 h in FLT3/ITD (BaF3/ITD, MOLM-13) and FLT3-WT (BaF3, HL-60) cells. N = 3 replicates. Bars, means ± S.D. Ctrl control cells treated with vehicle (DMSO), Orlis orlistat (10 μM), Quizar quizartinib (10 nM), Fato fatostatin (5 μM). C , D Western blot analysis showing knockdown of SREBP1 and FASN protein in BaF3/ITD and MOLM-13 cells by shRNA. NC non-specific shRNA sequence. E , F Quantitative analysis of cell death in BaF3/ITD and MOLM-13 cells following quizartinib treatment, with or without SREBP and FASN knockdown by shRNA.

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Control, Western Blot, Knockdown, shRNA, Sequencing

Journal: Cell Death & Disease

Article Title: Targeting oncogenic activation of FLT3/SREBP/FASN promotes the therapeutic effect of quizartinib involving disruption of mitochondrial phospholipids

doi: 10.1038/s41419-025-07661-6

Figure Lengend Snippet: A Illustration of the animal experiment: 2 days after tail vein injection of BaF3/ITD-EGFP-LUC2 cells, BALB/c mice were randomized and treated with fatostatin (15 mg/kg i.p. daily), orlistat (240 mg/kg i.p. daily), quizartinib (1 mg/kg oral daily), or quizartinib plus fatostatin/orlistat for another 28 days. B , C Leukemia burden and progression was monitored by luminescence imaging every 4 days at early stage and every week at late stage. n = 3 mice per group. D , E Survival was estimated by Kaplan–Meier analysis as described in Methods. n = 11 mice in each group. F , G Measurement of body weights of each group. Blank normal mice without leukemia cell injection, Ctrl control group with leukemia burden, Orlis orlistat, Fato fatostatin, Quizar quizartinib.

Article Snippet: The treatment of quizartinib and fatostatin was as the following: (1) Vehicle control (PBS containing 10% DMSO and 10% castor-oil); (2) 15 mg/kg fatostatin by intraperitoneal injection (dissolved in PBS containing 10% DMSO and 10% castor-oil); (3) 1 mg/kg quizartinib by oral gavage (dissolved in ddH 2 O) (MedChemExpress, Monmouth Junction, NJ, USA); (4) fatostatin + quizartinib.

Techniques: Injection, Imaging, Control