title: Distinct responses of compartmentalized glutathione redox potentials to pharmacologic quinones targeting NQO1.
abstract: Deoxynyboquinone (DNQ), a potent novel quinone-based antineoplastic agent, selectively kills solid cancers with overexpressed cytosolic NAD(P)H:quinone oxidoreductase-1 (NQO1) via excessive ROS production. A genetically encoded redox-sensitive probe was used to monitor intraorganellar glutathione redox potentials (EGSH) as a direct indicator of cellular oxidative stress following chemotherapeutic administration. Beta-lapachone (β-lap) and DNQ-induced spatiotemporal redox responses were monitored in human lung A549 and pancreatic MIA-PaCa-2 adenocarcinoma cells incubated with or without dicumarol and ES936, potent NQO1 inhibitors. Immediate oxidation of EGSHin both the cytosol and mitochondrial matrix was observed in response to DNQ and β-lap. The DNQ-induced cytosolic oxidation was fully prevented with NQO1 inhibition, whereas mitochondrial oxidation in A549 was NQO1-independent in contrast to MIA-PaCa-2 cells. However, at pharmacologic concentrations of β-lap both quinone-based substrates directly oxidized the redox probe, a possible sign of off-target reactivity with cellular thiols. Together, these data provide new evidence that DNQ's direct and discerning NQO1 substrate specificity underlies its pharmacologic potency, while β-lap elicits off-target responses at its effective doses.
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States. Electronic address: viadimer@illinois.edu.
authors.forename: Vladimir L
authors.initials: VL
authors.lastname: Kolossov
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Nagendraprabhu
authors.initials: N
authors.lastname: Ponnuraj
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Jessica N
authors.initials: JN
authors.lastname: Beaudoin
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Leslie
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Kenis
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: H Rex
authors.initials: HR
authors.lastname: Gaskins
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D001728:Dicumarol / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000032:analysis; D054477:Glutaredoxins / Q000032:analysis / Q000235:genetics; D005978:Glutathione / Q000032:analysis / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D045563:Indolequinones / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D057054:Molecular Imaging; D015335:Molecular Probes / Q000235:genetics; D058990:Molecular Targeted Therapy; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009285:Naphthoquinones / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D011809:Quinones / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: C444919:5-methoxy-1,2-dimethyl-3-((4-nitrophenoxy)methyl)indole-4,7-dione; D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D054477:Glutaredoxins; D045563:Indolequinones; D015335:Molecular Probes; D009285:Naphthoquinones; D011809:Quinones; D017382:Reactive Oxygen Species; C550580:deoxynyboquinone; D049452:Green Fluorescent Proteins; C014638:beta-lapachone; D001728:Dicumarol; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D005978:Glutathione
keywords: DNQ and β-lapachone; Lung and pancreatic cancer; NQO1; Redox homeostasis; roGFP2 redox probe