title: Identifying a resistance determinant for the antimitotic natural products disorazole C1 and A1.
abstract: Disorazoles are macrocyclic polyketides first isolated from the fermentation broth of the myxobacterium Sorangium cellulosum. Both the major fermentation product disorazole A(1) and its much rarer companion disorazole C(1) exhibit potent cytotoxic activity against many human tumor cells. Furthermore, the disorazoles appear to bind tubulin uniquely among known antimitotic agents, promoting apoptosis or premature senescence. It is uncertain what conveys tumor cell sensitivity to these complex natural products. Therefore, we generated and characterized human tumor cells resistant to disorazole C(1). Resistant cells proved exceedingly difficult to generate and required single step mutagenesis with chronic stepwise exposure to increasing concentrations of disorazole C(1). Compared with wild-type HeLa cells, disorazole C(1)-resistant HeLa/DZR cells were 34- and 8-fold resistant to disorazole C(1) and disorazole A(1) growth inhibition, respectively. HeLa/DZR cells were also remarkably cross-resistant to vinblastine (280-fold), paclitaxel (2400-fold), and doxorubicin (47-fold) but not cisplatin, suggesting a multidrug-resistant phenotype. Supporting this hypothesis, MCF7/MDR cells were 10-fold cross-resistant to disorazole C(1). HeLa/DZR disorazole resistance was not durable in the absence of chronic compound exposure. Verapamil reversed HeLa/DZR resistance to disorazole C(1) and disorazole A(1). Moreover, HeLa/DZR cells expressed elevated levels of the drug resistance ATP-binding cassette ABCB1 transporter. Loss of ABCB1 by incubation with short interfering RNA restored sensitivity to the disorazoles. Thus, the multidrug resistance transporter ABCB1 can affect the cytotoxicity of both disorazole C(1) and A(1). Disorazole C(1), however, retained activity against cells resistant against the clinically used microtubule-stabilizing agent epothilone B.
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh Drug Discovery Institute, Biomedical Science Tower-3, Suite 10040, 3501 Fifth Ave., University of Pittsburgh, Pittsburgh, PA 15260, USA. lazo@pitt.edu
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
authors.forename: Celeste E
authors.initials: CE
authors.lastname: Reese
authors.forename: Andreas
authors.initials: A
authors.lastname: Vogt
authors.forename: Laura L
authors.initials: LL
authors.lastname: Vollmer
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Kitchens
authors.forename: Eckhard
authors.initials: E
authors.lastname: Günther
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Graham
authors.forename: Chad D
authors.initials: CD
authors.lastname: Hopkins
authors.forename: Peter
authors.initials: P
authors.lastname: Wipf
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000235:genetics; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D034261:Epothilones / Q000494:pharmacology; D006801:Humans; D018942:Macrolides / Q000494:pharmacology*; D010080:Oxazoles / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics; D050257:Tubulin Modulators / Q000494:pharmacology*
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000903:Antibiotics, Antineoplastic; D034261:Epothilones; D018942:Macrolides; D010080:Oxazoles; D034741:RNA, Small Interfering; D050257:Tubulin Modulators; C508682:disorazole A1; C436383:disorazole C1; C093788:epothilone B