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Chang, Qing

Chen, Ruidong

Hanash, Samir

Hou, Pingping

Huang, Ailing

Jain, Abhinav K

Katayama, Hiroyuki

Lan, Zhengdao

Liang, Xin

Lin, Sue-Hwa

Logothetis, Christopher J., MD

Ma, Xingdi

Mak, Celia Sze Ling

Park, Jiwon

Peng, Ruiqing

Song, Jian H

Tang, Ming

Wang, Guocan

Zheng, Shui-Er

Zhu, Ming

English

OpenWorks @ MD Anderson

References1) Calses et al. Trends Cancer. 2019;5:297-307.2) Ehsanian et al. Oncogene. 2010;29:6160-71.3) Yuan et al. Cell Death Differ. 2018;15:1752-9.4) Yu et al. Clin Cancer Res. 2013;19:1389-99.5) Matallanas et al. Mol Cell. 20007;27:962-75.6) Cottini et al. Nat Med. 2014;20:599-606.7) Levy et al. Cell Death Differ. 2007;14:743-51.8) Cheung et al. Cell Stem Cell. 2020;27:590-604.9) Ou et al. Oncotarget. 2017;8:75727-41.10) Cheng  et al. Prostate Cancer Prostatic Dis.11) Nguyen et al. Cancer Cell. 2015;27:797–808.12) Wang et al. Cancer Discov. 2016;6:80-95.13) Zhang et al. Mol Cell Biol. 2015;35:1350–62.14) Ming et al. Oncogene. 2021, 40: 6049-65.Proposed Model for Tumor Suppressive Role of YAP1 in MetastasisBFigure 10. Proposed Model for Tumor Suppressive Role of YAP1in Metastasis. A model for hydroxylation-dependent YAP1 functionin cell migration, invasion, and metastasis (Created withBioRender.com). Left: P4HA2-mediated hydroxylation of YAP1 mayimpair its interactions with transcription factors such as JUN orenhance the recruitment of corepressor, such as SWI/SNF-NCoR1,NuRD, and EZH2/YY1, which results in a decrease in the expressionof genes involved in cell migration, invasion, and metastasis. Right:In the absence of P4HA2, non-hydroxylated YAP1 may efficientlyinteract with transcription factors such as JUN to activate genesinvolved in cell migration, invasion, and metastasis.YAP1 Deletion/mutation is Associated with Poor Survival and MetastasisFigure 1. Yap1 deletion/mutation is associated with poorsurvival and metastasis. (A) Disease free (Left) and Overall(Right) survival curve of patients with Yap1 WT ordeletion/mutation. (B) Percentage of primary and metastasistumors in patients with Yap1 WT or deletion/mutation.ABFigure 8. P4HA2 suppresses cell migration/invasion. (A) WBanalysis of P4HA2 in P4ha2-WT and P4ha2-KO PS cells. (B, C)Cell migration and invasion using P4ha2-WT and P4ha2-KO PScells. (D) qPCR analysis of YAP1 target genes (Postn, Col12a1,and Mgp) in P4ha2 KO and WT cells.Yap1 Hydroxylation Suppress Prostate Cancer MetastasisMing Zhu1, Ruiqing Peng1, 2, Xin Liang1, Zhengdao Lan3, Ming Tang4, Pingping Hou3, Jian H Song1, Celia Sze Ling Mak1, Jiwon Park1, Shui-Er Zheng1, Ailing Huang1, Xingdi Ma3, Ruidong Chen1, Qing Chang4, Christopher J Logothetis1, Abhinav K Jain5, Sue-Hwa Lin1, 6, Hiroyuki Katayama7, Samir Hanash7, Guocan Wang1, 81 Department of Genitourinary Medical Oncology and the David H. Koch Center for Applied Research of Genitourinary Cancers, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 2 Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China. 3 Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 4 Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 5 Department of Epigenetics and Molecular Carcinogenesis & Epigenomics Profiling Core Facility, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 6 Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 7 Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 8 Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. C Figure 5. YAP1 is hydroxylated at multiple proline residues.Multiple prolyl hydroxylation sites were identified in peptides ofYAP1 isoform 3 from the LC- MS/MS analysis.AbstractYes-associated protein 1 (YAP1), a key transcriptionalcoactivator in the Hippo pathway, is an importantdriver in cancer development and progression (1). Itplays an oncogenic role in various cancer types.However, multiple studies also support a tumor-suppressive function for YAP1 (2-8). Thus, thefunctions of YAP1 are likely context-dependent (9).On the cellular level, YAP1 promotes prostate cancercell proliferation (10-13). Yet, the role of YAP1 inprostate cancer cell invasion, migration, andmetastasis is not well defined. Through functional,transcriptomic, epigenomic, and proteomic analyses,we showed that prolyl hydroxylation of YAP1 plays acritical role in the suppression of cell migration,invasion, and metastasis in prostate cancer (14).Knockdown (KD) or knockout (KO) of YAP1 led toan increase in cell migration, invasion, and metastasisin prostate cancer cells. Microarray analysis showedthat the EMT pathway was activated in Yap1-KDcells. ChIP-seq analysis showed that YAP1 targetgenes are enriched in pathways regulating cellmigration. Mass spectrometry analysis identified P4Hprolyl hydroxylase in the YAP1 complex and YAP1was hydroxylated at multiple proline residues.Proline-to-alanine mutations of YAP1 isoform 3identified proline 174 as a critical residue, and itshydroxylation suppressed cell migration, invasion,and metastasis. KO of P4ha2 led to an increase in cellmigration and invasion, which was reversed uponYap1 KD. Our study identified a novel regulatorymechanism of YAP1 by which P4HA2-dependentprolyl hydroxylation of YAP1 determines itstranscriptional activities and its function in prostatecancer metastasis.ResultsA YAP1 Hydroxylation-defective Mut4-5 (PA) Promotes Cell Migration/invasion and MetastasisYAP1 Suppresses Cell Migration/Invasion and MetastasisBCCAFigure 2. Yap1 suppresses cell migration/invasion andmetastasis. (A, B) Cell migration and invasion assay usingPten/Smad4-deficieint prostate cancer (PS) cells transducedwith control shRNA and Yap1 shRNAs. The Yap1 KD efficiencywas confirmed by WB analysis. (C) Luciferase imaging in miceinjected with PS cells transduced with control shRNA and Yap1shRNAs through the tail vein.YAP1 Suppresses Pathways Involved in Cell Mobility and MetastasisFigure 3. YAP1 suppresses pathways involved in cellmobility and metastasis. (A) GSEA analysis of microarray datafrom PS cells transduced with doxycycline-inducible Yap1shRNA identified EMT as the top pathway activated in Yap1-KDcells. (B, C) ChIP-seq analysis identified YAP1 binding sites andYAP1-regulated pathways.E A D C Figure 5. YAP1 hydroxylation-defective Mut4-5 (PA)promotes cell migration/invasion and metastasis. (A)Scheme showing the strategy to generate prolyl hydroxylation–defective YAP1 mutants by mutating proline to alanine (PA):Mut1-3 (P75/85/120A), Mut4-5 (P172/174A), and Mut6-9(P348/352/394/397). Human YAP1 isoform 3 was used. (B)Expression of YAP1 WT and PA mutants in Yap1-KO PS cells.(C, D) Cell migration and invasion assay in Yap1-KO PS cellswith overexpression of YAP1 WT and PA mutants. (E) Tail veininjection of Yap1-KO cells with overexpression of YAP1 WT andPA mutants.A P4HA2 Suppresses Cell Migration/invasionYAP1 is Hydroxylated at Multiple ProlineResiduesB YAP1 Interacts with P4HA2Figure 4. YAP1 Interacts with P4HA2. (A) Immunoprecipitation-mass spectrometry analysis identified known YAP1-interactingproteins and novel YAP1-interacting proteins. (B) ExogenousYAP1 interacts with exogenous P4HA2 when overexpressed in293T cells by transfection of the indicated plasmids for co-immunoprecipitation experiments. (C) Endogenous YAP1interacts with endogenous P4HA2 in PS cells.B A B YAP1 P174 is Critical for Suppressing Cell Migration/invasionC Figure 7. YAP1 P174 is Critical for Suppressing CellMigration/invasion. (A, B) Cell migration and invasion assay usingYap1-KO PS cells with overexpression of GFP, YAP1 WT, Mut4-5, Mut4,and Mut5. (C) ChIP-qPCR analysis of YAP1, H3K4me3, and H3K9me4binding sites in Col12a1 and Mgp.A B C DP4HA2 Suppresses Cell Migration/invasion through YAP1Figure 9. P4HA2 Suppresses Cell Migration/invasion throughYAP1. (A) WB analysis of YAP1 in P4ha2-WT and P4ha2-KO PScells transduced with Yap1 shRNAs. (B) Cell invasion assay inP4ha2-WT and P4ha2-KO PS cells transduced with shYap1#434.BA

Yap1 Hydroxylation Suppress Prostate Cancer Metastasis

https://openworks.mdanderson.org/cgi/viewcontent.cgi?article=1031&context=leading-edge

Yap1 Hydroxylation Suppress Prostate Cancer Metastasis

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