bi4916  (MedChemExpress)

Journal: Frontiers in Oncology

Article Title: Dual targeting of glutamine and serine metabolism in acute myeloid leukemia

doi: 10.3389/fonc.2024.1326754

Figure Lengend Snippet: AML cells sensitive to PHGDH inhibition by small molecule inhibitor BI4916. (A) Approximately 18 h after plating, MOLM-14, U937, MV4-11 and MonoMac-6 cell lines were treated for 72 h with increasing doses of BI4916. Proliferation of cells was measured by the addition of WST-1. IC 50 s were generated by GraphPad Prism. The table shows BI4916 mean IC 50 values (µM) ± standard deviation (SD) for the AML cell lines tested. (B) MOLM-14 and U937 cells were treated with fixed doses of BI4916 and Rylaze for 72 h. After WST-1 termination, combination indexes (CI) were calculated using Compusyn Software, using Chou Talalay’s method. CI values were calculated using Compusyn. (C) MOLM-14 and U937 cells were treated with BI4916 (2 µM), Rylaze (0.1 µg/mL), and BI4916-Rylaze treatment for 72 h and the percentage of cell death was measured using trypan blue exclusion, data is expressed as % cytotoxicity (n=3). (D) MOLM14 and U937 cells were treated as in (C) and GSH was measured by a luminescence-based assay 72 h post-treatment. Data is expressed as the percentage of GSH relative to vehicle control (n=3). MOLM-14 (E) and U937 (F) cells were pre-loaded with H 2 DCFA, a cell permeable indicator of ROS, then treated as in (C) or 200 µM of H 2 O 2 as a positive control. ROS was measured at 0, 1, 2, 4, 6, and 24 h by plate reader. Results were normalized to control and expressed as mean ± standard error of the mean (SEM) (n=3). (G) MOLM-14 and U937 cell lines were treated as in (C) and glutamate levels were measured by a luminescence-based assay 72 h post treatment. Data is expressed as the percentage of Relative Light units (RLU) relative to vehicle control (n=3). ns, not significant, *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001.

Article Snippet: BI4916 (Cat#: HY-126253) and GCN2-IN-1 (Cat#: HY-100877) was purchased from MedChem Express (Monmouth Junction, NJ) as a powder and dissolved in DMSO in 50 mM stock solutions and stored at -20°C.

Techniques: Inhibition, Generated, Standard Deviation, Software, Luminescence Assay, Control, Positive Control

Journal: Frontiers in Oncology

Article Title: Dual targeting of glutamine and serine metabolism in acute myeloid leukemia

doi: 10.3389/fonc.2024.1326754

Figure Lengend Snippet: Combination of BI4916 and Rylaze demonstrates synergism and potent anti-proliferative effect on primary human AML samples. (A) Clinical course, cytogenetics and molecular mutations of patient-derived primary AML cells. (B) Primary AML cells were plated overnight and then treated with fixed ratios of BI4916 (called BI in the figure) and Rylaze (called Ryl in the figure) alone and in combination. Cultures were terminated at 48 h and viability was assessed with AlamarBlue. CI values were plotted with their respective combination ratios. (C) Healthy BMMC plated overnight then treated with increasing doses of BI4916, Rylaze or the combination. Cultures were terminated at 48 h and viability was assessed with Alamar Blue (n=2).

Article Snippet: BI4916 (Cat#: HY-126253) and GCN2-IN-1 (Cat#: HY-100877) was purchased from MedChem Express (Monmouth Junction, NJ) as a powder and dissolved in DMSO in 50 mM stock solutions and stored at -20°C.

Techniques: Derivative Assay

Journal: Frontiers in Oncology

Article Title: Dual targeting of glutamine and serine metabolism in acute myeloid leukemia

doi: 10.3389/fonc.2024.1326754

Figure Lengend Snippet: BI4916 and Rylaze impedes cap-dependent translation and protein synthesis. (A) MOLM-14 and U937 were treated with BI4916 (2 µM), Rylaze (0.1 µg/mL) and BI4916-Rylaze for 16 h followed by 20 min of incubation with puromycin (1 µg/mL) and then lysed. Cell lysates were subjected to immunoblotting with the anti-puromycin antibody (SUnSET [surface sensing of translation] assay). The bar diagram represents densitometric quantification of 3 independent experiments. (B) MOLM-14 and U937 cells were treated as in (A) , and cellular lysates were probed with the indicated antibodies. Bar diagram represents densitometric quantification of experiments (n=3). (C) Cellular lysates of MOLM-14 and U937 treated as in (A) were incubated with m 7 GTP sepharose beads for 2 h, followed by immunoblotting and probing with the indicated antibodies. The bar diagram represents densitometric quantification (n = 3). (D) Schematic of proposed mechanism of BI4916-Rylaze treatment on cap-dependent mRNA translation made with BioRender. ns, not significant, *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001.

Article Snippet: BI4916 (Cat#: HY-126253) and GCN2-IN-1 (Cat#: HY-100877) was purchased from MedChem Express (Monmouth Junction, NJ) as a powder and dissolved in DMSO in 50 mM stock solutions and stored at -20°C.

Techniques: Incubation, Western Blot

Journal: Frontiers in Oncology

Article Title: Dual targeting of glutamine and serine metabolism in acute myeloid leukemia

doi: 10.3389/fonc.2024.1326754

Figure Lengend Snippet: Schematic of Rylaze and BI4916 effect on AML cells. Phosphoglycerate Dehydrogenase (PHGDH) regulates de novo serine biosynthesis using glycolytic intermediate 3-phosphoglycerate (3-PG). An oxidative reaction converts 3-PG to 3-phosphohydroxypyruvate which leads to de novo serine biosynthesis. Targeting PHGDH with BI4916 inhibits de novo serine biosynthesis which synergizes with glutamine deprivation mediated by Rylaze (crisantaspase). BI4916-Rylaze combination leads to decreased glutathione production and protein synthesis and increased cell death.

Article Snippet: BI4916 (Cat#: HY-126253) and GCN2-IN-1 (Cat#: HY-100877) was purchased from MedChem Express (Monmouth Junction, NJ) as a powder and dissolved in DMSO in 50 mM stock solutions and stored at -20°C.

Techniques: