title: Inhibition of LRRK2 kinase activity stimulates macroautophagy.
abstract: Leucine Rich Repeat Kinase 2 (LRRK2) is one of the most important genetic contributors to Parkinson's disease. LRRK2 has been implicated in a number of cellular processes, including macroautophagy. To test whether LRRK2 has a role in regulating autophagy, a specific inhibitor of the kinase activity of LRRK2 was applied to human neuroglioma cells and downstream readouts of autophagy examined. The resulting data demonstrate that inhibition of LRRK2 kinase activity stimulates macroautophagy in the absence of any alteration in the translational targets of mTORC1, suggesting that LRRK2 regulates autophagic vesicle formation independent of canonical mTORC1 signaling. This study represents the first pharmacological dissection of the role LRRK2 plays in the autophagy/lysosomal pathway, emphasizing the importance of this pathway as a marker for LRRK2 physiological function. Moreover it highlights the need to dissect autophagy and lysosomal activities in the context of LRRK2 related pathologies with the final aim of understanding their aetiology and identifying specific targets for disease modifying therapies in patients.
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK. Electronic address: c.manzoni@ucl.ac.uk.
authors.forename: Claudia
authors.initials: C
authors.lastname: Manzoni
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK; Reta Lila Weston Institute and Queen Square Brain Bank, UCL Institute of Neurology, 1 Wakefield Street, London, WC1N 1PJ, UK.
authors.forename: Adamantios
authors.initials: A
authors.lastname: Mamais
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
authors.forename: Sybille
authors.initials: S
authors.lastname: Dihanich
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
authors.forename: Rosella
authors.initials: R
authors.lastname: Abeti
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
authors.forename: Marc P M
authors.initials: MPM
authors.lastname: Soutar
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
authors.forename: Helene
authors.initials: H
authors.lastname: Plun-Favreau
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
authors.forename: Paola
authors.initials: P
authors.lastname: Giunti
authors.affiliation: London Research Institute, Cancer Research UK, Lincoln's Inn Fields, London, WC2A 3LY, UK.
authors.forename: Sharon A
authors.initials: SA
authors.lastname: Tooze
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK; Reta Lila Weston Institute and Queen Square Brain Bank, UCL Institute of Neurology, 1 Wakefield Street, London, WC1N 1PJ, UK.
authors.forename: Rina
authors.initials: R
authors.lastname: Bandopadhyay
authors.affiliation: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK; School of Pharmacy, University of Reading, Whiteknights, Reading, RG6 6AP, UK. Electronic address: patrick.lewis@ucl.ac.uk.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Lewis
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D001343:Autophagy / Q000187:drug effects; D001570:Benzodiazepinones / Q000494:pharmacology; D045744:Cell Line, Tumor; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D008869:Microtubule-Associated Proteins; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D000071456:Sequestosome-1 Protein
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D001570:Benzodiazepinones; C582847:LRRK2-IN1; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; C495280:LRRK2 protein, human; C547428:LRRK2 protein, rat; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases
keywords: C-terminal of ROC domain; COR; DMSO; DPBS; Dimethylsulfoxide; Dulbecco's phosphate buffered saline; EDTA; Ethylene di-ammonium tetra acetic acid; LC3; LRRK2; Leucine Rich Repeat Kinase 2; Macroautophagy; Mammalian target of rapamycin; Parkinson's disease; ROC; Ras of Complex Proteins; SDS; Sodium dodecyl sulphate; WIPI2; mTOR; p62
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