title: A mechanistic study of tumor-targeted corrole toxicity.
abstract: HerGa is a self-assembled tumor-targeted particle that bears both tumor detection and elimination activities in a single, two-component complex (Agadjanian et al. Proc. Natl. Acad. Sci. U.S.A.2009, 106, 6105-6110). Given its multifunctionality, HerGa (composed of the fluorescent cytotoxic corrole macrocycle, S2Ga, noncovalently bound to the tumor-targeted cell penetration protein, HerPBK10) has the potential for high clinical impact, but its mechanism of cell killing remains to be elucidated, and hence is the focus of the present study. Here we show that HerGa requires HerPBK10-mediated cell entry to induce toxicity. HerGa (but not HerPBK10 or S2Ga alone) induced mitochondrial membrane potential disruption and superoxide elevation, which were both prevented by endosomolytic-deficient mutants, indicating that cytosolic exposure is necessary for corrole-mediated cell death. A novel property discovered here is that corrole fluorescence lifetime acts as a pH indicator, broadcasting the intracellular microenvironmental pH during uptake in live cells. This feature in combination with two-photon imaging shows that HerGa undergoes early endosome escape during uptake, avoiding compartments of pH < 6.5. Cytoskeletal disruption accompanied HerGa-mediated mitochondrial changes whereas oxygen scavenging reduced both events. Paclitaxel treatment indicated that HerGa uptake requires dynamic microtubules. Unexpectedly, low pH is insufficient to induce release of the corrole from HerPBK10. Altogether, these studies identify a mechanistic pathway in which early endosomal escape enables HerGa-induced superoxide generation leading to cytoskeletal and mitochondrial damage, thus triggering downstream cell death.
authors: Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, United States.
authors: Jae Youn
authors: JY
authors: Hwang
authors: Jay
authors: J
authors: Lubow
authors: David
authors: D
authors: Chu
authors: Jun
authors: J
authors: Ma
authors: Hasmik
authors: H
authors: Agadjanian
authors: Jessica
authors: J
authors: Sims
authors: Harry B
authors: HB
authors: Gray
authors: Zeev
authors: Z
authors: Gross
authors: Daniel L
authors: DL
authors: Farkas
authors: Lali K
authors: LK
authors: Medina-Kauwe
mesh_terms: D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems; D006801:Humans; D006863:Hydrogen-Ion Concentration; D000922:Immunotoxins / Q000633:toxicity*; D018613:Microscopy, Confocal; D008954:Models, Biological; D009369:Neoplasms / Q000473:pathology; D017239:Paclitaxel / Q000494:pharmacology; D011166:Porphyrins / Q000633:toxicity*
chemical_list: D000922:Immunotoxins; D011166:Porphyrins; C431863:corrole; D017239:Paclitaxel