Abstract
Acute promyelocytic leukemia (APL) is driven by the oncoprotein PML/RARα, which destroys the architecture of PML nuclear bodies (NBs). PML NBs are critical to tumor suppression, and their disruption mediated by PML/RARα accelerates APL pathogenesis. However, the mechanisms of PML NB disruption remain elusive. Here, we reveal that the failure of NB assembly in APL results from neddylation-induced aberrant phase separation of PML/RARα. Mechanistically, PML/RARα is neddylated in the RARα moiety, and this neddylation enhances its DNA-binding ability and further impedes the phase separation of the PML moiety, consequently disrupting PML NB construction. Accordingly, deneddylation of PML/RARα restores its phase separation process to reconstruct functional NBs and activates RARα signaling, thereby suppressing PML/RARα-driven leukemogenesis. Pharmacological inhibition of neddylation by MLN4924 eradicates APL cells both in vitro and in vivo. Our work elucidates the neddylation-destroyed phase separation mechanism for PML/RARα-driven NB disruption and highlights targeting neddylation for APL eradication.
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Data availability
Raw data for RNA-sequencing are deposited at the NCBI Gene Expression Omnibus (GSE165046). Other data is available in the main text or the supplementary materials. For more additional data related to this subject, please contact the corresponding author.
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Acknowledgements
Dr. Lingtao Wu (University of Southern California) kindly donated NB4 cell line. NB4R1 cells were kindly gifted from Dr. He Huang (Zhejiang University) and NB4R2 cells were kindly provided by Dr. Jian Zhang (Shanghai Jiao Tong University). OCI-AML2 cells were kindly gifted from Dr. Yu Rao (Tsinghua Univesity).
Funding
This work was supported by grants from the National Natural Science Foundation of China (Nos. 81973354 to MY, No. 81803552 to XS); Leading Talent of “Ten Thousand Plan”—National High-Level Talents Special Support Plan and the Fundamental Research Funds for the Central Universities.
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MY and XS conceived the study and analyzed data; XS performed high-content screening; YC performed phase separation and BioID assay; XS and AX performed the mHSPC-related experiment; XS, YC, AX, DX, WD and YH performed the immunofluorescence and western blotting procedure; AX, WW, MC and ZX detected the therapeutic effect of MLN4924; WW generated the PML KO cells; WW and XZ quantified the immunofluorescence data; JC, QH, BY, YW and YZ conceived the experiments and helped organize the paper; MY, XS and YC wrote the manuscript.
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Shao, X., Chen, Y., Xu, A. et al. Deneddylation of PML/RARα reconstructs functional PML nuclear bodies via orchestrating phase separation to eradicate APL. Cell Death Differ 29, 1654–1668 (2022). https://doi.org/10.1038/s41418-022-00955-8
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DOI: https://doi.org/10.1038/s41418-022-00955-8
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