Curcumin and resveratrol sensitizes TP53 mutant acute myeloid leukemia (AML) cell lines to venetoclax-induced apoptosis

Abstract While venetoclax-based regimens have significantly improved outcomes for many patients with acute myeloid leukemia (AML), those with the TP53 mutation often remain resistant, posing a treatment challenge. Curcumin and resveratrol, two natural compounds, have been shown to restore TP53 function in other TP53 mutant malignancies. We hypothesized that Curcumin and resveratrol would restore TP53 function, and subsequently, restore sensitivity to venetoclax to induce apoptosis in TP53-mutant AML. Three TP53-mutant cell lines; HL-60, MOLM-13, and KG-1, were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum for 72 hours. Cells were seeded at a 20,000 cells/well in 96-well plates in triplicate with escalating concentrations of venetoclax [0.01 nM-100 µM] either alone or in combination with curcumin [0.1µM-100µM] and/or resveratrol [0.1µ-100µM]. Following treatment, cells were incubated for an additional 48 hours. Apoptosis was quantified using a Caspase-7 bioluminescence assay, with data normalized to the untreated control. A two-way ANOVA evaluated treatment effects at 100 µM venetoclax. Additionally, for each cell line and venetoclax concentration, a one-way ANOVA compared the effects of curcumin, resveratrol, and their combination, followed by post-hoc Tukey HSD tests. Normality and variance homogeneity were verified using Shapiro-Wilk and Levene's tests. At 100 µM venetoclax, the combination of curcumin and resveratrol (both at 100 µM) significantly enhanced apoptosis compared to venetoclax alone across all three cell lines—HL-60 (p < 0.01), MOLM-13 (p < 0.01), and KG-1 (p = 0.03)—with apoptotic means ranging from 41.49-101.82 (for the combination) versus 30.06-90.05 (venetoclax alone) versus 33.12-83.67 (curcumin alone) versus 33.55-88.49 (resveratrol alone); indicating a synergistic interaction. Curcumin alone outperformed resveratrol at 100 µM in MOLM13 (88.49 vs. 83.67, p < 0.05) and KG1 (5.03 vs. 3.76, p < 0.01), a trend that persisted at 10 µM venetoclax (3.42 vs. 1.98, p < 0.05) and without venetoclax (2.26 vs. 1.07, p < 0.05) in KG-1. Optimal synergy occurred at 100 µM venetoclax, particularly in MOLM13 (101.82 vs. 83.67-90.05). In the absence of venetoclax, the one-way ANOVA showed no significant differences in HL60 and MOLM13 (p > 0.05). However, with 0 µM venetoclax in KG1, curcumin 100 µM and curcumin + resveratrol; both at 100 µM, induced increased apoptosis, compared to no additives (p < 0.01). Cell line-specific responses were significant (p < 0.05), with MOLM13 showing the greatest venetoclax-induced apoptosis and KG1 demonstrating sensitivity to high-dose curcumin alone. To conclude, combined use of curcumin and resveratrol enhanced venetoclax-induced apoptosis in vitro, in TP53 mutant cell lines with notable synergy observed in MOLM-13. Curcumin alone also showed promising effects in the KG-1 cell line. These findings suggest these natural compounds to be potential candidates for adjunct therapies in TP53-mutant AML and may warrant future evaluation. Follow up experiments in primary TP53-AML mutant cell samples and assays of TP53 function are currently ongoing.