title: The LRRK2 inhibitor GSK2578215A induces protective autophagy in SH-SY5Y cells: involvement of Drp-1-mediated mitochondrial fission and mitochondrial-derived ROS signaling.
abstract: Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene have been associated with Parkinson's disease, and its inhibition opens potential new therapeutic options. Among the drug inhibitors of both wild-type and mutant LRRK2 forms is the 2-arylmethyloxy-5-subtitutent-N-arylbenzamide GSK257815A. Using the well-established dopaminergic cell culture model SH-SY5Y, we have investigated the effects of GSK2578215A on crucial neurodegenerative features such as mitochondrial dynamics and autophagy. GSK2578215A induces mitochondrial fragmentation of an early step preceding autophagy. This increase in autophagosome results from inhibition of fusion rather than increases in synthesis. The observed effects were shared with LRRK2-IN-1, a well-described, structurally distinct kinase inhibitor compound or when knocking down LRRK2 expression using siRNA. Studies using the drug mitochondrial division inhibitor 1 indicated that translocation of the dynamin-related protein-1 has a relevant role in this process. In addition, autophagic inhibitors revealed the participation of autophagy as a cytoprotective response by removing damaged mitochondria. GSK2578215A induced oxidative stress as evidenced by the accumulation of 4-hydroxy-2-nonenal in SH-SY5Y cells. The mitochondrial-targeted reactive oxygen species scavenger MitoQ positioned these species as second messengers between mitochondrial morphologic alterations and autophagy. Altogether, our results demonstrated the relevance of LRRK2 in mitochondrial-activated pathways mediating in autophagy and cell fate, crucial features in neurodegenerative diseases.
authors.affiliation: 1] Grupo de Neurofarmacología, Dpto. Ciencias Médicas, Facultad de Medicina de Albacete, Universidad de Castilla-La Mancha, IDINE, Albacete, Spain [2] Facultad de Medicina y Odontología, Universidad Católica de Valencia 'San Vicente Mártir' Valencia, Valencia, Spain [3] Unidad de Neuropsicofarmacología Traslacional, Complejo Hospitalario Universitario de Albacete, Albacete, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Saez-Atienzar
authors.affiliation: Facultad de Medicina y Odontología, Universidad Católica de Valencia 'San Vicente Mártir' Valencia, Valencia, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Bonet-Ponce
authors.affiliation: Facultad de Medicina y Odontología, Universidad Católica de Valencia 'San Vicente Mártir' Valencia, Valencia, Spain.
authors.forename: J R
authors.initials: JR
authors.lastname: Blesa
authors.affiliation: Facultad de Medicina y Odontología, Universidad Católica de Valencia 'San Vicente Mártir' Valencia, Valencia, Spain.
authors.forename: F J
authors.initials: FJ
authors.lastname: Romero
authors.affiliation: Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge, UK.
authors.forename: M P
authors.initials: MP
authors.lastname: Murphy
authors.affiliation: Grupo de Neurofarmacología, Dpto. Ciencias Médicas, Facultad de Medicina de Albacete, Universidad de Castilla-La Mancha, IDINE, Albacete, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Jordan
authors.affiliation: Unidad de Neuropsicofarmacología Traslacional, Complejo Hospitalario Universitario de Albacete, Albacete, Spain.
authors.forename: M F
authors.initials: MF
authors.lastname: Galindo
mesh_terms: D000447:Aldehydes / Q000378:metabolism; D000631:Aminopyridines / Q000633:toxicity*; D001343:Autophagy / Q000187:drug effects*; D001549:Benzamides / Q000633:toxicity*; D001570:Benzodiazepinones / Q000494:pharmacology; D045744:Cell Line, Tumor; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D063154:Mitochondrial Dynamics / Q000187:drug effects*; D024101:Mitochondrial Proteins / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000447:Aldehydes; D000631:Aminopyridines; D001549:Benzamides; D001570:Benzodiazepinones; C000600930:GSK2578215A; C582847:LRRK2-IN1; D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D009943:Organophosphorus Compounds; D011743:Pyrimidines; D034741:RNA, Small Interfering; D014451:Ubiquinone; C429014:mitoquinone; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins; C027576:4-hydroxy-2-nonenal