title: Identification of 12/15-lipoxygenase as a regulator of axon degeneration through high-content screening.
abstract: Axon degeneration is a programed process that takes place during development, in response to neuronal injury, and as a component of neurodegenerative disease pathology, yet the molecular mechanisms that drive this process remain poorly defined. In this study, we have developed a semi-automated, 384-well format axon degeneration assay in rat dorsal root ganglion (DRG) neurons using a trophic factor withdrawal paradigm. Using this setup, we have screened a library of known drugs and bioactives to identify several previously unappreciated regulators of axon degeneration, including lipoxygenases. Multiple structurally distinct lipoxygenase inhibitors as well as mouse DRG neurons lacking expression of 12/15-lipoxygenase display protection of axons in this context. Retinal ganglion cell axons from 12/15-lipoxygenase-null mice were similarly protected from degeneration following nerve crush injury. Through additional mechanistic studies, we demonstrate that lipoxygenases act cell autonomously within neurons to regulate degeneration, and are required for mitochondrial permeabilization and caspase activation in the axon. These findings suggest that these enzymes may represent an attractive target for treatment of neuropathies and provide a potential mechanism for the neuroprotection observed in various settings following lipoxygenase inhibitor treatment.
authors.affiliation: In Vitro Pharmacology, Evotec AG, Manfred Eigen Campus, 22419 Hamburg, Germany, lewcock.joseph@gene.com york.rudhard@evotec.com.
authors.forename: York
authors.initials: Y
authors.lastname: Rudhard
authors.affiliation: Departments of Neuroscience.
authors.forename: Arundhati
authors.initials: A
authors.lastname: Sengupta Ghosh
authors.affiliation: In Vitro Pharmacology, Evotec AG, Manfred Eigen Campus, 22419 Hamburg, Germany.
authors.forename: Beatrix
authors.initials: B
authors.lastname: Lippert
authors.affiliation: In Vitro Pharmacology, Evotec AG, Manfred Eigen Campus, 22419 Hamburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Böcker
authors.affiliation: In Vitro Pharmacology, Evotec AG, Manfred Eigen Campus, 22419 Hamburg, Germany.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Pedaran
authors.affiliation: In Vitro Pharmacology, Evotec AG, Manfred Eigen Campus, 22419 Hamburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Krämer
authors.affiliation: Pathology, and.
authors.forename: Hai
authors.initials: H
authors.lastname: Ngu
authors.affiliation: Pathology, and.
authors.forename: Oded
authors.initials: O
authors.lastname: Foreman
authors.affiliation: Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California 94080.
authors.forename: Yichin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Departments of Neuroscience, lewcock.joseph@gene.com york.rudhard@evotec.com.
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Lewcock
mesh_terms: D000465:Algorithms; D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001369:Axons / Q000378:metabolism / Q000473:pathology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D015723:Gene Library; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009410:Nerve Degeneration / Q000175:diagnosis / Q000188:drug therapy / Q000201:enzymology* / Q000209:etiology; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009901:Optic Nerve Diseases / Q000150:complications; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
chemical_list: C475064:12-15-lipoxygenase; D004791:Enzyme Inhibitors; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase
keywords: DRG; NGF; axon degeneration; lipoxygenase