title: Ligand-independent androgen receptor variants derived from splicing of cryptic exons signify hormone-refractory prostate cancer.
abstract: Suppression of androgen production and function provides palliation but not cure in men with prostate cancer (PCa). Therapeutic failure and progression to hormone-refractory PCa (HRPC) are often accompanied by molecular alterations involving the androgen receptor (AR). In this study, we report novel forms of AR alteration that are prevalent in HRPC. Through in silico sequence analysis and subsequent experimental validation studies, we uncovered seven AR variant transcripts lacking the reading frames for the ligand-binding domain due to splicing of "intronic" cryptic exons to the upstream exons encoding the AR DNA-binding domain. We focused on the two most abundantly expressed variants, AR-V1 and AR-V7, for more detailed analysis. AR-V1 and AR-V7 mRNA showed an average 20-fold higher expression in HRPC (n = 25) when compared with hormone-naive PCa (n = 82; P < 0.0001). Among the hormone-naive PCa, higher expression of AR-V7 predicted biochemical recurrence following surgical treatment (P = 0.012). Polyclonal antibodies specific to AR-V7 detected the AR-V7 protein frequently in HRPC specimens but rarely in hormone-naive PCa specimens. AR-V7 was localized in the nuclei of cultured PCa cells under androgen-depleted conditions, and constitutively active in driving the expression of canonical androgen-responsive genes, as revealed by both AR reporter assays and expression microarray analysis. These results suggest a novel mechanism for the development of HRPC that warrants further investigation. In addition, as expression markers for lethal PCa, these novel AR variants may be explored as potential biomarkers and therapeutic targets for advanced PCa.
authors.affiliation: Department of Urology, Oncology, and Pathology, The Johns Hopkins University School of Medicine, 600 North Wolfe Street, Baltimore, MD 21287, USA.
authors.forename: Rong
authors.initials: R
authors.lastname: Hu
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Dunn
authors.forename: Shuanzeng
authors.initials: S
authors.lastname: Wei
authors.forename: Sumit
authors.initials: S
authors.lastname: Isharwal
authors.forename: Robert W
authors.initials: RW
authors.lastname: Veltri
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Humphreys
authors.forename: Misop
authors.initials: M
authors.lastname: Han
authors.forename: Alan W
authors.initials: AW
authors.lastname: Partin
authors.forename: Robert L
authors.initials: RL
authors.lastname: Vessella
authors.forename: William B
authors.initials: WB
authors.lastname: Isaacs
authors.forename: G Steven
authors.initials: GS
authors.lastname: Bova
authors.forename: Jun
authors.initials: J
authors.lastname: Luo
mesh_terms: D017398:Alternative Splicing; D003001:Cloning, Molecular; D005091:Exons; D006801:Humans; D008024:Ligands; D008297:Male; D009376:Neoplasms, Hormone-Dependent / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D011944:Receptors, Androgen / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D008024:Ligands; D020033:Protein Isoforms; D011944:Receptors, Androgen