title: HIF prolyl hydroxylase inhibition increases cell viability and potentiates dopamine release in dopaminergic cells.
abstract: Hypoxia-inducible factor (HIF) controls the expression of genes that adapts the cellular condition to accommodate oxidative stress. The potential beneficial effect of HIF up-regulation in ischemia has recently gained interest substantiated by the known HIF-regulation of erythropoietin and other hypoxia accommodating genes. So far the perspectives for HIF up-regulation has been focused on anemia and ischemia related diseases but little information is available about the relevance of HIF biology for neurodegenerative disease like Parkinson's disease. We therefore sought out to characterize the effect of HIF-up-regulation on survival and dopamine homeostasis in dopaminergic cells. We used a low molecular weight HIF prolyl hydroxylase (HPH) inhibitor and lentiviral based shRNA knockdown of HPH subtypes as molecular tools to increase HIF protein level and downstream HIF-regulated genes. We show that HIF induction results in protection against oxidative stress in cellular models based on PC12 cells and LUHMES cells. In addition, HPH inhibition elevates tyrosine hydroxylase expression and activity, which causes increased dopamine synthesis and release in both PC12 cells and a primary rat ventral mesencephalic cell culture. All together these findings suggest that prolyl hydroxylases may represent novel targets for therapeutic intervention in disorders characterized by dopamine homeostasis dysregulation like Parkinson's disease.
authors.affiliation: Discovery Biology Research, H. Lundbeck A/S, Valby, Denmark.
authors.forename: Jens Leander
authors.initials: JL
authors.lastname: Johansen
authors.forename: Thomas Nikolaj
authors.initials: TN
authors.lastname: Sager
authors.forename: Julie
authors.initials: J
authors.lastname: Lotharius
authors.forename: Louise
authors.initials: L
authors.lastname: Witten
authors.forename: Arne
authors.initials: A
authors.lastname: Mørk
authors.forename: Jan
authors.initials: J
authors.lastname: Egebjerg
authors.forename: Kenneth
authors.initials: K
authors.lastname: Thirstrup
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D049308:Dioxygenases / Q000037:antagonists & inhibitors*; D004298:Dopamine / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D016716:PC12 Cells; D011393:Procollagen-Proline Dioxygenase / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D049308:Dioxygenases; D008156:Luciferases; C552717:HIF prolyl hydroxylase, rat; D011393:Procollagen-Proline Dioxygenase; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine