title: Acetylcholinesterase inhibitors increase ADAM10 activity by promoting its trafficking in neuroblastoma cell lines.
abstract: Acetylcholinesterase inhibitors (AChEIs) are the only currently available drugs for treating Alzheimer's Disease (AD). Some authors have suggested a function of AChEIs not only in the induction of AChE overproduction and alternative splicing shifts but also a possible role of these drugs in amyloid metabolism beyond their well-known symptomatic effect. Here, we investigate the mechanisms of action of the AChEI donepezil on APP (amyloid precursor protein) metabolism and on the activity/trafficking of the alpha-secretase candidate ADAM 10, in differentiated human neuroblastoma cells (SH-SY5Y). In these cells, the activity of AChE is significantly decreased after 2 h of donepezil treatment. Further, SH-SY5Y cells released significantly more sAPPalpha into the medium, whereas total APP levels in cell lysates were unchanged. Interestingly, treated cells showed increased ADAM 10 levels in membrane compartments. This effect was prevented by pretreatment with tunicamycin or brefeldin, suggesting that donepezil affects trafficking and/or maturation of ADAM 10; additionally, this pretreatment significantly decreased sAPPalpha levels. Pre-incubation with atropine decreased release of sAPPalpha significantly but did not revert ADAM 10 activity to control levels further suggesting that donepezil acts not solely through a purely receptor mediated pathway. These findings indicate that donepezil exerts multiple mechanisms involving processing and trafficking of key proteins involved in AD pathogenesis.
authors.affiliation: Centre of Excellence on Neurodegenerative Diseases and Department of Pharmacological Sciences, University of Milan, Milan, Italy. Martina.Zimmermann@unimi.it
authors.forename: Martina
authors.initials: M
authors.lastname: Zimmermann
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Gardoni
authors.forename: Elena
authors.initials: E
authors.lastname: Marcello
authors.forename: Francesca
authors.initials: F
authors.lastname: Colciaghi
authors.forename: Barbara
authors.initials: B
authors.lastname: Borroni
authors.forename: Alessandro
authors.initials: A
authors.lastname: Padovani
authors.forename: Flaminio
authors.initials: F
authors.lastname: Cattabeni
authors.forename: Monica
authors.initials: M
authors.lastname: Di Luca
mesh_terms: D051722:ADAM Proteins; D000072197:ADAM10 Protein; D000110:Acetylcholinesterase / Q000378:metabolism; D053829:Amyloid Precursor Protein Secretases; D016229:Amyloid beta-Peptides / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D001285:Atropine / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D020126:Brefeldin A / Q000494:pharmacology; D002038:Bungarotoxins / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002458:Cell Fractionation / Q000379:methods; D045744:Cell Line, Tumor; D018679:Cholinergic Agonists / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D000077265:Donepezil; D004347:Drug Interactions; D004622:Embryo, Mammalian; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D007189:Indans / Q000494:pharmacology*; D008565:Membrane Proteins / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000379:methods; D018727:Muscarinic Antagonists / Q000494:pharmacology; D009447:Neuroblastoma / Q000473:pathology; D010830:Physostigmine / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D002038:Bungarotoxins; D018679:Cholinergic Agonists; D002800:Cholinesterase Inhibitors; D007189:Indans; D008565:Membrane Proteins; D018727:Muscarinic Antagonists; D010880:Piperidines; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; D014212:Tretinoin; D001285:Atropine; D000077265:Donepezil; D002217:Carbachol; D010830:Physostigmine; D000110:Acetylcholinesterase; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human