Wu-5, a novel USP10 inhibitor, enhances crenolanib-induced FLT3-ITD-positive AML cell death via inhibiting FLT3 and AMPK pathways

  • Acta Pharmacol Sin. 2021 Apr;42(4):604-612. doi: 10.1038/s41401-020-0455-x.
Miao Yu  #  1  2 ,  Zhi-Xiao Fang  #  2 ,  Wei-Wei Wang  2 ,  Ying Zhang  1  2 ,  Zhi-Lei Bu  1  2 ,  Meng Liu  2 ,  Xin-Hua Xiao  3 ,  Zi-Lu Zhang  2  3 ,  Xing-Ming Zhang  2  4 ,  Yang Cao  5 ,  Ying-Ying Wang  2 ,  Hu Lei  2 ,  Han-Zhang Xu  2 ,  Yun-Zhao Wu  2 ,  Wei Liu  6 ,  Ying-Li Wu  7
Affiliations
  • 1. Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
  • 2. Hongqiao International Institute of Medicine, Shanghai Tongren Hospital/Faculty of Basic MedicineChemical Biology Division of Shanghai Universities E-Institutes, Key Laboratory of Cell Differentiation and Apoptosis of the Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
  • 3. Department of Hematology, Shanghai Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
  • 4. Department of Transfusion Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
  • 5. Department of Hematology, The Third Affiliated Hospital of Soochow University, The First People's Hospital of Changzhou, Changzhou, 213003, China.
  • 6. Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China. [email protected].
  • 7. Hongqiao International Institute of Medicine, Shanghai Tongren Hospital/Faculty of Basic MedicineChemical Biology Division of Shanghai Universities E-Institutes, Key Laboratory of Cell Differentiation and Apoptosis of the Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China. [email protected].
  • # Contributed equally.
PMID: 32694757 DOI: 10.1038/s41401-020-0455-x
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.

Keywords
AML; AMPKα; Compound C; FLT3-ITD; USP10; Wu-5; crenolanib; metformin.
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