Blockade of deubiquitinase YOD1 degrades oncogenic PML/RAR α and eradicates acute promyelocytic leukemia cells

Xuejing Shao; Yingqian Chen; Wei Wang; Wenxin Du; Xingya Zhang; Minyi Cai; Shaowei Bing; Ji Cao; Xiaojun Xu; Bo Yang; Qiaojun He; Meidan Ying

doi:10.1016/j.apsb.2021.10.020

Blockade of deubiquitinase YOD1 degrades oncogenic PML/RAR α and eradicates acute promyelocytic leukemia cells

Acta Pharm Sin B. 2022 Apr;12(4):1856-1870. doi: 10.1016/j.apsb.2021.10.020. Epub 2021 Oct 23.

Authors

Xuejing Shao¹, Yingqian Chen¹, Wei Wang¹, Wenxin Du¹, Xingya Zhang¹, Minyi Cai¹, Shaowei Bing¹, Ji Cao^{1

2

3}, Xiaojun Xu⁴, Bo Yang^{1

3}, Qiaojun He^{1

2

3}, Meidan Ying^{1

4

2}

Affiliations

¹ Institute of Pharmacology and Toxicology, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
² Cancer Center, Zhejiang University, Hangzhou 310058, China.
³ Innovation Institute for Artificial Intelligence in Medicine, Zhejiang University, Hangzhou 310058, China.
⁴ Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou 310052, China.

Abstract

In most acute promyelocytic leukemia (APL) cells, promyelocytic leukemia (PML) fuses to retinoic acid receptor α (RARα) due to chromosomal translocation, thus generating PML/RARα oncoprotein, which is a relatively stable oncoprotein for degradation in APL. Elucidating the mechanism regulating the stability of PML/RARα may help to degrade PML/RARα and eradicate APL cells. Here, we describe a deubiquitinase (DUB)-involved regulatory mechanism for the maintenance of PML/RARα stability and develop a novel pharmacological approach to degrading PML/RARα by inhibiting DUB. We utilized a DUB siRNA library to identify the ovarian tumor protease (OTU) family member deubiquitinase YOD1 as a critical DUB of PML/RARα. Suppression of YOD1 promoted the degradation of PML/RARα, thus inhibiting APL cells and prolonging the survival time of APL cell-bearing mice. Subsequent phenotypic screening of small molecules allowed us to identify ubiquitin isopeptidase inhibitor I (G5) as the first YOD1 pharmacological inhibitor. As expected, G5 notably degraded PML/RARα protein and eradicated APL, particularly drug-resistant APL cells. Importantly, G5 also showed a strong killing effect on primary patient-derived APL blasts. Overall, our study not only reveals the DUB-involved regulatory mechanism on PML/RARα stability and validates YOD1 as a potential therapeutic target for APL, but also identifies G5 as a YOD1 inhibitor and a promising candidate for APL, particularly drug-resistant APL treatment.

Keywords: APL, acute promyelocytic leukemia; ATO, arsenic trioxide; ATRA, all-trans retinoic acid; Acute promyelocytic leukemia; Degradation; Deubiquitinase; Drug resistance; EARD, endoplasmic reticulum-associated degradation; FLT3/ITD, internal tandem duplication within FLT3; G5, ubiquitin isopeptidase inhibitor I; HOTAIRM1, HOXA transcript antisense RNA myeloid-specific 1; Inhibitor; JAMM, Jab1/Pab1/MPN domain-containing protease; LATS, large tumor suppressor kinase; MDM2, murine double minute 2; MINDY, motif-interacting with ubiquitin-containing novel DUB family; MJD, machado-Joseph domain-containing protease; OUT, ovarian tumor; PLZF, promyelocytic leukemia zinc finger; PML, promyelocytic leukemia; PML/RARα; RARα, retinoic acid receptor α; RNF4, ring finger protein 4; S100A3, S100 calcium binding protein A3; TAZ, transcriptional co-activator with PDZ-binding motif; TGFβ, transforming growth factor β; TRIB3, tribbles pseudokinase 3; Therapy; UCH, ubiquitin carboxyl terminal hydrolase; UCHL1, ubiquitin c-terminal hydrolase L1; USP, ubiquitin specific protease; YAP, yes-associated protein; YOD1; cAMP, cyclic adenosine monophosphate.