fig6

Figure 6. In vitro and in vivo validation of the synergistic anti-AML effects of Elesclomol and Venetoclax. (A-C) Dose-response curves and matrices for Venetoclax and ES-Cu in OCI-AML3 (ZIP synergy score: 14.371), U937 (ZIP synergy score: 11.321), and MOLM13/Vene (ZIP synergy score: 12.915) cells, with treatment for 24 h before viability measurement using CCK-8; (D-F) Apoptosis of the AML cell lines in Cluster 3 (U937, OCI-AML3,MOLM13/VENE) detected by flow cytometry after treatment with 100 nM ES-Cu, 3 μM Venetoclax, and a combination of ES-Cu and Venetoclax; (G) Apoptosis of primary cells from refractory AML patients detected by flow cytometry after treatment with 100 nM ES-Cu, 1 μM Venetoclax, and a combination of ES-Cu and Venetoclax; (H) Expression levels of apoptotic protein caspase-3 and its cleaved form in primary AML patient samples following treatment with ES-Cu, Venetoclax, and their combination; (I) Dose-response curves and matrices for Venetoclax and ES-Cu in stem cell from healthy donor (ZIP synergy score: 3.058); (J) Establishment of the Venetoclax-resistant mouse model and the corresponding dosing regimen; (K) Spleen morphology and weight in mice after combination treatment with Elesclomol (10 mg/kg) and Venetoclax (50 mg/kg); (L and M) Tumor burden (hCD45⁺ cells) in the bone marrow (L) and spleen (M) of mice following combination treatment with Elesclomol and Venetoclax. Results are expressed as mean ± SD of at least three independent experiments. Statistical significance was determined using an unpaired Student’s t-test. P < 0.05 was considered statistically significant. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. AML: Acute myeloid leukemia; ES-Cu: elesclomol-copper; ZIP: zero interaction potency; CCK-8: Cell Counting Kit-8; SD: standard deviation; IC₅₀: half-maximal inhibitory concentration; APC: allophycocyanin; SSC: side scatter.