Abstract
The kinase FLT3 internal tandem duplication (FLT3-ITD) is related to poor clinical outcomes of acute myeloid leukemia (AML). FLT3 inhibitors have provided novel strategies for the treatment of FLT3-ITD-positive AML. But they are limited by rapid development of acquired resistance and refractory in monotherapy. Recent evidence shows that inducing the degradation of FLT3-mutated protein is an attractive strategy for the treatment of FLT3-ITD-positive AML, especially those with FLT3 inhibitor resistance. In this study we identified Wu-5 as a novel USP10 inhibitor inducing the degradation of FLT3-mutated protein. We showed that Wu-5 selectively inhibited the viability of FLT3 inhibitor-sensitive (MV4-11, Molm13) and -resistant (MV4-11R) FLT3-ITD-positive AML cells with IC50 of 3.794, 5.056, and 8.386 μM, respectively. Wu-5 (1−10 μM) dose-dependently induced apoptosis of MV4-11, Molm13, and MV4-11R cells through the proteasome-mediated degradation of FLT3-ITD. We further demonstrated that Wu-5 directly interacted with and inactivated USP10, the deubiquitinase for FLT3-ITD in vitro (IC50 value = 8.3 µM) and in FLT3-ITD-positive AML cells. Overexpression of USP10 abrogated Wu-5-induced FLT3-ITD degradation and cell death. Also, the combined treatment of Wu-5 and crenolanib produced synergistic cell death in FLT3-ITD-positive cells via the reduction of both FLT3 and AMPKα proteins. In support of this, AMPKα inhibitor compound C synergistically enhanced the anti-leukemia effect of crenolanib, while AMPKα activator metformin inhibited the anti-leukemia effect of crenolanib. In summary, we demonstrate that Wu-5, a novel USP10 inhibitor, can overcome FLT3 inhibitor resistance and synergistically enhance the anti-AML effect of crenolanib through targeting FLT3 and AMPKα pathway.
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Acknowledgements
This work was supported in part by grants from the National Key Research and Development Program of China (No. 2017YFA0505202), the National Basic Research Program of China (973 Program) (No. 2015CB910403), the National Natural Science Foundation of China (Nos. 81570118, 81700475, 81570112, 81670139, and 21602133), the National Natural Science Foundation Youths of China (No. 81700157), Innovative research team of high-level local universities in Shanghai (SSMU-ZDCX20181202).
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YLW and WL conceived and designed the study. MY and ZXF performed the experiments, collected data, conducted preliminary analysis of data and wrote the initial article. WWW, YZ, ZLB, ML, XHX, ZLZ, XMZ, YC, YYW, HL, HZX and YZW assisted in data analysis, manuscript writing and editing. All authors reviewed the manuscript.
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Yu, M., Fang, Zx., Wang, Ww. et al. Wu-5, a novel USP10 inhibitor, enhances crenolanib-induced FLT3-ITD-positive AML cell death via inhibiting FLT3 and AMPK pathways. Acta Pharmacol Sin 42, 604–612 (2021). https://doi.org/10.1038/s41401-020-0455-x
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