title: Mis-trafficking of endosomal urokinase proteins triggers drug-induced glioma nonapoptotic cell death.
abstract: 5-Benzylglycinyl-amiloride (UCD38B) is the parent molecule of a class of anticancer small molecules that kill proliferative and nonproliferative high-grade glioma cells by programmed necrosis. UCD38B intracellularly triggers endocytosis, causing 40-50% of endosomes containing proteins of the urokinase plasminogen activator system (uPAS) to relocate to perinuclear mitochondrial regions. Endosomal "mis-trafficking" caused by UCD38B in human glioma cells corresponds to mitochondrial depolarization with the release and nuclear translocation of apoptotis-inducing factor (AIF) followed by irreversible caspase-independent cell demise. High-content quantification of immunocytochemical colocalization studies identified that UCD38B treatment increased endocytosis of the urokinase plasminogen activator (uPA), its receptor (uPAR), and plasminogen activator inhibitor-1 (PAI-1) into the early and late endosomes by 4- to 5-fold prior to AIF nuclear translocation and subsequent glioma demise. PAI-1 was found to comparably relocate with a subset of early and late endosomes in four different human glioma cell lines after UCD38B treatment, followed by caspase-independent, nonapoptotic cell death. Following UCD38B treatment, the receptor guidance protein LRP-1, which is required for endosomal recycling of the uPA receptor to the plasmalemma, remained abnormally associated with PAI-1 in early and late endosomes. The resultant aberrant endosomal recycling increased the total cellular content of the uPA-PAI-1 protein complex. Reversible inhibition of cellular endocytosis demonstrated that UCD38B bypasses the plasmalemmal uPAS complex and directly acts intracellularly to alter uPAS endocytotic trafficking. UCD38B represents a class of small molecules whose anticancer cytotoxicity is a consequence of causing the mis-trafficking of early and late endosomes containing uPAS cargo and leading to AIF-mediated necrotic cell death.
authors: Department of Neurology, School of Medicine (N.P., F.G.), and Department of Molecular Biosciences, School of Veterinary Medicine (N.P., A.C.G., F.G.), University of California, Davis, California fagorin@ucdavis.edu npasupul@ucdavis.edu.
authors: Nagarekha
authors: N
authors: Pasupuleti
authors: Department of Neurology, School of Medicine (N.P., F.G.), and Department of Molecular Biosciences, School of Veterinary Medicine (N.P., A.C.G., F.G.), University of California, Davis, California.
authors: Ana Cristina
authors: AC
authors: Grodzki
authors: Department of Neurology, School of Medicine (N.P., F.G.), and Department of Molecular Biosciences, School of Veterinary Medicine (N.P., A.C.G., F.G.), University of California, Davis, California fagorin@ucdavis.edu npasupul@ucdavis.edu.
authors: Fredric
authors: F
authors: Gorin
mesh_terms: D000584:Amiloride / Q000031:analogs & derivatives* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004705:Endocytosis / Q000187:drug effects; D011992:Endosomes / Q000378:metabolism; D005910:Glioma / Q000201:enzymology* / Q000473:pathology; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009336:Necrosis; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D021381:Protein Transport; D055293:Receptors, Urokinase Plasminogen Activator / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: C584412:5-benzylglycinyl amiloride; D000970:Antineoplastic Agents; D051033:Apoptosis Inducing Factor; C475476:LRP1 protein, human; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D017395:Plasminogen Activator Inhibitor 1; D055293:Receptors, Urokinase Plasminogen Activator; C051321:SERPINE1 protein, human; D000584:Amiloride; D014568:Urokinase-Type Plasminogen Activator; D005998:Glycine
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