title: Target sequencing and CRISPR/Cas editing reveal simultaneous loss of UTX and UTY in urothelial bladder cancer.
abstract: UTX is a histone demethylase gene located on the X chromosome and is a frequently mutated gene in urothelial bladder cancer (UBC). UTY is a paralog of UTX located on the Y chromosome. We performed target capture sequencing on 128 genes in 40 non-metastatic UBC patients. UTX was the most frequently mutated gene (30%, 12/40). Of the genetic alterations identified, 75% were truncating mutations. UTY copy number loss was detected in 8 male patients (22.8%, 8/35). Of the 9 male patients with UTX mutations, 6 also had copy number loss (66.7%). To evaluate the functional roles of UTX and UTY in tumor progression, we designed UTX and UTY single knockout and UTX-UTY double knockout experiments using a CRISPR/Cas9 lentiviral system, and compared the proliferative capacities of two UBC cell lines in vitro. Single UTX or UTY knockout increased cell proliferation as compared to UTX-UTY wild-type cells. UTX-UTY double knockout cells exhibited greater proliferation than single knockout cells. These findings suggest both UTX and UTY function as dose-dependent suppressors of UBC development. While UTX escapes X chromosome inactivation in females, UTY may function as a male homologue of UTX, which could compensate for dosage imbalances.
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, Korea.
authors.forename: Jinwoo
authors.initials: J
authors.lastname: Ahn
authors.affiliation: Department of Urology, Ewha Womans University School of Medicine, Seoul, Korea.
authors.forename: Kwang Hyun
authors.initials: KH
authors.lastname: Kim
authors.affiliation: Department of Pathology, Ewha Womans University School of Medicine, Seoul, Korea.
authors.forename: Sanghui
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Molecular Medicine, Ewha Womans University School of Medicine, Seoul, Korea.
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Ahn
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, Korea.
authors.forename: Ha Young
authors.initials: HY
authors.lastname: Kim
authors.affiliation: Department of Urology, Ewha Womans University School of Medicine, Seoul, Korea.
authors.forename: Hana
authors.initials: H
authors.lastname: Yoon
authors.affiliation: Department of Clinical Pharmacology and Therapeutics, College of Medicine, Kyung Hee University, Seoul, Korea.
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, Korea.
authors.forename: Duhee
authors.initials: D
authors.lastname: Bang
authors.affiliation: Department of Urology, Ewha Womans University School of Medicine, Seoul, Korea.
authors.forename: Dong Hyeon
authors.initials: DH
authors.lastname: Lee
mesh_terms: D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002843:Chromatin / Q000737:chemistry; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D044127:Epigenesis, Genetic; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D056466:Histone Demethylases / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008297:Male; D015778:Minor Histocompatibility Antigens / Q000378:metabolism*; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D011446:Prospective Studies; D001749:Urinary Bladder Neoplasms / Q000378:metabolism*; D049951:X Chromosome Inactivation
chemical_list: D002843:Chromatin; D015778:Minor Histocompatibility Antigens; D009687:Nuclear Proteins; C484453:UTY protein, human; D056466:Histone Demethylases; C523998:KDM6A protein, human
keywords: UTX; UTY; chromatin remodeling; epigenesis; urinary bladder neoplasm