title: Epigenetic silencing of argininosuccinate synthetase confers resistance to platinum-induced cell death but collateral sensitivity to arginine auxotrophy in ovarian cancer.
abstract: Evidence indicates that acquired resistance of cancers to chemotherapeutic agents can occur via epigenetic mechanisms. Down-regulation of expression of argininosuccinate synthetase (ASS1), the rate-limiting enzyme in the biosynthesis of arginine, has been associated with the development of platinum resistance in ovarian cancer treated with platinum-based chemotherapy. The aim of the present study was to analyse epigenetic regulation of ASS1 in ovarian cancer tissue taken at diagnosis and relapse and determine its significance as a predictor of clinical outcome in patients treated with platinum-based chemotherapy. In addition, expression and epigenetic regulation of ASS1 were analysed in human ovarian cancer cell lines, and ASS1 expression correlated with the ability of the lines to grow in media containing cisplatin, carboplatin or taxol or in arginine-depleted media. Our results show that aberrant methylation in the ASS1 promoter correlated with transcriptional silencing in ovarian cancer cell lines. ASS1 silencing conferred selective resistance to platinum-based drugs and conferred arginine auxotrophy and sensitivity to arginine deprivation. In ovarian cancer, ASS1 methylation at diagnosis was associated with significantly reduced overall survival (p = 0.01) and relapse-free survival (p = 0.01). In patients who relapse, ASS1 methylation was significantly more frequent at relapse (p = 0.008). These data establish epigenetic inactivation of ASS1 as a determinant of response to platinum chemotherapy and imply that transcriptional silencing of ASS1 contributes to treatment failure and clinical relapse in ovarian cancer. The collateral sensitivity of cells lacking endogenous ASS1 to arginine depletion suggests novel therapeutic strategies for the management of relapsed ovarian cancer.
authors.affiliation: King's College London School of Medicine, Cancer Studies Division, The Rayne Institute, St. Thomas' Hospital, London, United Kingdom. linda.nicholson@kcl.ac.uk
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Nicholson
authors.forename: Paul R
authors.initials: PR
authors.lastname: Smith
authors.forename: Louise
authors.initials: L
authors.lastname: Hiller
authors.forename: Peter W
authors.initials: PW
authors.lastname: Szlosarek
authors.forename: Christopher
authors.initials: C
authors.lastname: Kimberley
authors.forename: Jalid
authors.initials: J
authors.lastname: Sehouli
authors.forename: Dominique
authors.initials: D
authors.lastname: Koensgen
authors.forename: Alexander
authors.initials: A
authors.lastname: Mustea
authors.forename: Peter
authors.initials: P
authors.lastname: Schmid
authors.forename: Tim
authors.initials: T
authors.lastname: Crook
mesh_terms: D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D001120:Arginine / Q000172:deficiency*; D001124:Argininosuccinate Synthase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D016190:Carboplatin / Q000008:administration & dosage; D016923:Cell Death; D002945:Cisplatin / Q000008:administration & dosage; D018284:Cystadenocarcinoma, Serous / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D019175:DNA Methylation; D019008:Drug Resistance, Neoplasm; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D020868:Gene Silencing; D006801:Humans; D015151:Immunoblotting; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D017239:Paclitaxel / Q000008:administration & dosage; D011401:Promoter Regions, Genetic / Q000235:genetics; D015996:Survival Rate; D014407:Tumor Cells, Cultured
chemical_list: D001120:Arginine; D016190:Carboplatin; D001124:Argininosuccinate Synthase; D017239:Paclitaxel; D002945:Cisplatin