EZH2 crosstalk with RNA methylation promotes prostate cancer progression through modulation of m6A autoregulation pathway
Yang Yi, Joshua Fry, Chaehyun Yum, Rui Wang, Siqi Wu, Sharath Narayan, Qi Liu, Xingxing Zhang, Htoo Zarni Oo, Ning Xie, Yanqiang Li, Xinlei Gao, Xufen Yu, Xiaoping Hu, Qiaqia Li, Kemal Keseroglu, Ertuğrul M. Özbudak, Sarki A. Abdulkadir, Kaifu Chen, Jian Jin, Jonathan C. Zhao, Xuesen Dong, Daniel Arango, Rendong Yang, Qi Cao
Yang Yi, Joshua Fry, Chaehyun Yum, Rui Wang, Siqi Wu, Sharath Narayan, Qi Liu, Xingxing Zhang, Htoo Zarni Oo, Ning Xie, Yanqiang Li, Xinlei Gao, Xufen Yu, Xiaoping Hu, Qiaqia Li, Kemal Keseroglu, Ertuğrul M. Özbudak, Sarki A. Abdulkadir, Kaifu Chen, Jian Jin, Jonathan C. Zhao, Xuesen Dong, Daniel Arango, Rendong Yang, Qi Cao
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Research Article Cell biology Oncology

EZH2 crosstalk with RNA methylation promotes prostate cancer progression through modulation of m6A autoregulation pathway

Abstract

N6-methyladenosine (m6A), the most predominant RNA modification in humans, participates in various fundamental and pathological bioprocesses. Dynamic manipulation of m6A deposition in the transcriptome is critical for cancer progression, though how this regulation is achieved remains understudied. Here, we report that, in prostate cancer (PCa), Polycomb group (PcG) protein Enhancer of Zeste Homolog 2 (EZH2) exerts an additional function in m6A regulation via its enzymatic activity. Mechanistically, EZH2 methylates and stabilizes FOXA1 proteins from degradation, which, in turn, facilitates the transcription of m6A reader YTHDF1. Through activating an m6A autoregulation pathway, YTHDF1 enhances the translation of METTL14 and WTAP, 2 critical components of the m6A methyltransferase complex (MTC), and thereby upregulates the global m6A level in PCa cells. We further demonstrate that inhibiting the catalytic activity of EZH2 suppresses the translation process globally through targeting the YTHDF1-m6A axis. By disrupting both the expression and interaction of key m6A MTC subunits, combinational treatment of EZH2 degrader MS8815 and m6A inhibitor STM2457 mitigates prostate tumor growth synergistically. Together, our study decodes a previously hidden interrelationship between EZH2 and mRNA modification, which may be leveraged to advance the EZH2-targeting curative strategies in cancer.

Authors

Yang Yi, Joshua Fry, Chaehyun Yum, Rui Wang, Siqi Wu, Sharath Narayan, Qi Liu, Xingxing Zhang, Htoo Zarni Oo, Ning Xie, Yanqiang Li, Xinlei Gao, Xufen Yu, Xiaoping Hu, Qiaqia Li, Kemal Keseroglu, Ertuğrul M. Özbudak, Sarki A. Abdulkadir, Kaifu Chen, Jian Jin, Jonathan C. Zhao, Xuesen Dong, Daniel Arango, Rendong Yang, Qi Cao

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Figure 1

EZH2 maintains a hyper-m6A state in PCa cells.

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EZH2 maintains a hyper-m6A state in PCa cells. (A) The m6A ELISA to meas...
(A) The m6A ELISA to measure the global m6A levels in PrEC and BPH-1 benign prostate cells, along with a panel of PCa cell lines. (B) Western blot to detect the EZH2 expression level in all cell types tested in A. (C and D) The m6A ELISA to measure the global m6A levels in 2 PCa cell lines upon EZH2 knockdown (C), or in PrEC cells with EZH2 overexpression (D). (E) Representative fluorescence images to show the RNA m6A staining in control and EZH2-deficient C4-2 cells. Endogenous EZH2 were costained by anti-EZH2 antibody, and the nuclei were visualized by DAPI (Scale bar: 20 μm). (F) The m6A ELISA to measure the global m6A levels in C4-2 cells treated with a series of EZH2 inhibitors. For DZNeP, GSK126, and EPZ6438, a concentration of 5 μM was used. For all the MS drugs, a concentration of 1 μM was used. (G) Scatter plot showing the m6A methylation in C4-2 cells upon EZH2 depletion, with siEZH2 hypomethylated sites in blue and hypermethylated sites in red. (H) Violin plot to show the differential m6A ratio upon EZH2 knockdown in C4-2 cells. (I) The distribution of EZH2-affected m6A sites across the transcript. (J) Gene Ontology enrichment analysis of genes with hypomethylated m6A sites upon EZH2 knockdown in C4-2 cells. Statistical significance was assessed using the hypergeometric test with P values corrected for multiple testing with the g:SCS method using g:Profiler. (K) The m6A CUT&RUN-qPCR analysis to validate the EZH2-affected m6A sites in each indicated transcript of C4-2 cells. One-way ANOVA followed by Dunnett’s multiple-comparison test was used for statistical analysis in A, C, F, and K. Two-tailed Student’s t test was used in D.