title: The dual-acting chemotherapeutic agent Alchemix induces cell death independently of ATM and p53.
abstract: Topoisomerase inhibitors are in common use as chemotherapeutic agents although they can display reduced efficacy in chemotherapy-resistant tumours, which have inactivated DNA damage response (DDR) genes, such as ATM and TP53. Here, we characterise the cellular response to the dual-acting agent, Alchemix (ALX), which is a modified anthraquinone that functions as a topoisomerase inhibitor as well as an alkylating agent. We show that ALX induces a robust DDR at nano-molar concentrations and this is mediated primarily through ATR- and DNA-PK- but not ATM-dependent pathways, despite DNA double strand breaks being generated after prolonged exposure to the drug. Interestingly, exposure of epithelial tumour cell lines to ALX in vitro resulted in potent activation of the G2/M checkpoint, which after a prolonged arrest, was bypassed allowing cells to progress into mitosis where they ultimately died by mitotic catastrophe. We also observed effective killing of lymphoid tumour cell lines in vitro following exposure to ALX, although, in contrast, this tended to occur via activation of a p53-independent apoptotic pathway. Lastly, we validate the effectiveness of ALX as a chemotherapeutic agent in vivo by demonstrating its ability to cause a significant reduction in tumour cell growth, irrespective of TP53 status, using a mouse leukaemia xenograft model. Taken together, these data demonstrate that ALX, through its dual action as an alkylating agent and topoisomerase inhibitor, represents a novel anti-cancer agent that could be potentially used clinically to treat refractory or relapsed tumours, particularly those harbouring mutations in DDR genes.
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: A
authors.initials: A
authors.lastname: Thomas
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: T
authors.initials: T
authors.lastname: Perry
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: S
authors.initials: S
authors.lastname: Berhane
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: C
authors.initials: C
authors.lastname: Oldreive
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: A
authors.initials: A
authors.lastname: Zlatanou
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: L R
authors.initials: LR
authors.lastname: Williams
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: V J
authors.initials: VJ
authors.lastname: Weston
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: T
authors.initials: T
authors.lastname: Stankovic
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: P
authors.initials: P
authors.lastname: Kearns
authors.affiliation: Institute of Cancer Therapeutics, School of Life Sciences, University of Bradford, West Yorkshire, UK.
authors.forename: K
authors.initials: K
authors.lastname: Pors
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: R J
authors.initials: RJ
authors.lastname: Grand
authors.affiliation: IBR West Extension, School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham, UK.
authors.forename: G S
authors.initials: GS
authors.lastname: Stewart
mesh_terms: D000818:Animals; D000880:Anthraquinones / Q000494:pharmacology* / Q000627:therapeutic use; D000951:Antigens, Neoplasm / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004249:DNA Damage / Q000187:drug effects; D004260:DNA Repair / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D006801:Humans; D007945:Leukemia, Lymphoid / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D051379:Mice; D059003:Topoisomerase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
chemical_list: C483940:Alchemix; D000880:Anthraquinones; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; D059003:Topoisomerase Inhibitors; D016159:Tumor Suppressor Protein p53; D064007:Ataxia Telangiectasia Mutated Proteins; D004250:DNA Topoisomerases, Type II