title: Catalase abrogates β-lapachone-induced PARP1 hyperactivation-directed programmed necrosis in NQO1-positive breast cancers.
abstract: Improving patient outcome by personalized therapy involves a thorough understanding of an agent's mechanism of action. β-Lapachone (clinical forms, Arq501/Arq761) has been developed to exploit dramatic cancer-specific elevations in the phase II detoxifying enzyme NAD(P)H:quinone oxidoreductase (NQO1). NQO1 is dramatically elevated in solid cancers, including primary and metastatic [e.g., triple-negative (ER-, PR-, Her2/Neu-)] breast cancers. To define cellular factors that influence the efficacy of β-lapachone using knowledge of its mechanism of action, we confirmed that NQO1 was required for lethality and mediated a futile redox cycle where ∼120 moles of superoxide were formed per mole of β-lapachone in 2 minutes. β-Lapachone induced reactive oxygen species (ROS), stimulated DNA single-strand break-dependent poly(ADP-ribose) polymerase-1 (PARP1) hyperactivation, caused dramatic loss of essential nucleotides (NAD(+)/ATP), and elicited programmed necrosis in breast cancer cells. Although PARP1 hyperactivation and NQO1 expression were major determinants of β-lapachone-induced lethality, alterations in catalase expression, including treatment with exogenous enzyme, caused marked cytoprotection. Thus, catalase is an important resistance factor and highlights H2O2 as an obligate ROS for cell death from this agent. Exogenous superoxide dismutase enhanced catalase-induced cytoprotection. β-Lapachone-induced cell death included apoptosis-inducing factor (AIF) translocation from mitochondria to nuclei, TUNEL+ staining, atypical PARP1 cleavage, and glyceraldehyde 3-phosphate dehydrogenase S-nitrosylation, which were abrogated by catalase. We predict that the ratio of NQO1:catalase activities in breast cancer versus associated normal tissue are likely to be the major determinants affecting the therapeutic window of β-lapachone and other NQO1 bioactivatable drugs.
authors.affiliation: Corresponding Authors: Erik A. Bey, West Virginia University, 1 Medical Center Drive, Box 9300, Room 1835, Morgantown, WV 26506. ebey@hsc.wvu.edu.
authors.forename: Erik A
authors.initials: EA
authors.lastname: Bey
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Reinicke
authors.forename: Melissa C
authors.initials: MC
authors.lastname: Srougi
authors.forename: Marie
authors.initials: M
authors.lastname: Varnes
authors.forename: Vernon E
authors.initials: VE
authors.lastname: Anderson
authors.forename: John J
authors.initials: JJ
authors.lastname: Pink
authors.forename: Long Shan
authors.initials: LS
authors.lastname: Li
authors.forename: Malina
authors.initials: M
authors.lastname: Patel
authors.forename: Lifen
authors.initials: L
authors.lastname: Cao
authors.forename: Zachary
authors.initials: Z
authors.lastname: Moore
authors.forename: Amy
authors.initials: A
authors.lastname: Rommel
authors.forename: Michael
authors.initials: M
authors.lastname: Boatman
authors.forename: Cheryl
authors.initials: C
authors.lastname: Lewis
authors.forename: David M
authors.initials: DM
authors.lastname: Euhus
authors.forename: William G
authors.initials: WG
authors.lastname: Bornmann
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Buchsbaum
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Spitz
authors.forename: Jinming
authors.initials: J
authors.lastname: Gao
authors.forename: David A
authors.initials: DA
authors.lastname: Boothman
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D053904:DNA Breaks, Single-Stranded / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000235:genetics / Q000378:metabolism*; D009285:Naphthoquinones / Q000008:administration & dosage*; D009336:Necrosis / Q000235:genetics / Q000473:pathology; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D009285:Naphthoquinones; D017382:Reactive Oxygen Species; C014638:beta-lapachone; D006861:Hydrogen Peroxide; D002374:Catalase; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases