title: Manipulation of prostate cancer metastasis by locus-specific modification of the CRMP4 promoter region using chimeric TALE DNA methyltransferase and demethylase.
abstract: Prostate cancer is the most commonly diagnosed non-cutaneous cancer and one of the leading causes of cancer death for North American men. Whereas localized prostate cancer can be cured, there is currently no cure for metastatic prostate cancer. Here we report a novel approach that utilizes designed chimeric transcription activator-like effectors (dTALEs) to control prostate cancer metastasis. Transfection of dTALEs of DNA methyltransferase or demethylase induced artificial, yet active locus-specific CpG and subsequent histone modifications. These manipulations markedly altered expression of endogenous CRMP4, a metastasis suppressor gene. Remarkably, locus-specific CpG demethylation of the CRMP4 promoter in metastatic PC3 cells abolished metastasis, whereas locus-specific CpG methylation of the promoter in non-metastatic 22Rv1 cells induced metastasis. CRMP4-mediated metastasis suppression was found to require activation of Akt/Rac1 signaling and down-regulation of MMP-9 expression. This proof-of-concept study with dTALEs for locus-specific epigenomic manipulation validates the selected CpG methylation of CRMP4 gene as an independent biomarker for diagnosis and prognosis of prostate cancer metastasis and opens up a novel avenue for mechanistic research on cancer biology.
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Pang
authors.affiliation: Department of Pharmacology, Uniformed Services University of The Health Sciences, Bethesda, MD 20814, USA.
authors.forename: Huaiyan
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Department of Urology, The First Affiliated Hospital of Nan Chang University, Nanchang 330006, China.
authors.forename: Wei-Peng
authors.initials: WP
authors.lastname: Liu
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Jin-Ming
authors.initials: JM
authors.lastname: Di
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Heng-Jun
authors.initials: HJ
authors.lastname: Xiao
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Wen-Tao
authors.initials: WT
authors.lastname: Huang
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Ming-Kun
authors.initials: MK
authors.lastname: Chen
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Liao-Yuan
authors.initials: LY
authors.lastname: Li
authors.affiliation: Department of Pathology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Chun-Kui
authors.initials: CK
authors.lastname: Shao
authors.affiliation: Department of Pharmacology, Uniformed Services University of The Health Sciences, Bethesda, MD 20814, USA.
authors.forename: Ying-Hong
authors.initials: YH
authors.lastname: Feng
authors.affiliation: Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Gao
mesh_terms: D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009124:Muscle Proteins / Q000235:genetics*; D009362:Neoplasm Metastasis; D009419:Nerve Tissue Proteins / Q000235:genetics*; D011379:Prognosis; D011401:Promoter Regions, Genetic; D064129:Prostatic Neoplasms, Castration-Resistant / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D014162:Transfection
chemical_list: C526904:Dpysl3 protein, mouse; C102926:DPYSL3 protein, human; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D015254:DNA Modification Methylases
keywords: CRMP4; epigenetic manipulation; metastasis; prostate cancer; transcription activator-like effectors (TALEs)