title: Autophagic lysosome reformation dysfunction in glucocerebrosidase deficient cells: relevance to Parkinson disease.
abstract: Glucocerebrosidase (GBA1) gene mutations increase the risk of Parkinson disease (PD). While the cellular mechanisms associating GBA1 mutations and PD are unknown, loss of the glucocerebrosidase enzyme (GCase) activity, inhibition of autophagy and increased α-synuclein levels have been implicated. Here we show that autophagy lysosomal reformation (ALR) is compromised in cells lacking functional GCase. ALR is a cellular process controlled by mTOR which regenerates functional lysosomes from autolysosomes formed during macroautophagy. A decrease in phopho-S6K levels, a marker of mTOR activity, was observed in models of GCase deficiency, including primary mouse neurons and the PD patient derived fibroblasts with GBA1 mutations, suggesting that ALR is compromised. Importantly Rab7, a GTPase crucial for endosome-lysosome trafficking and ALR, accumulated in GCase deficient cells, supporting the notion that lysosomal recycling is impaired. Recombinant GCase treatment reversed ALR inhibition and lysosomal dysfunction. Moreover, ALR dysfunction was accompanied by impairment of macroautophagy and chaperone-mediated autophagy, increased levels of total and phosphorylated (S129) monomeric α-synuclein, evidence of amyloid oligomers and increased α-synuclein release. Concurrently, we found increased cholesterol and altered glucosylceramide homeostasis which could compromise ALR. We propose that GCase deficiency in PD inhibits lysosomal recycling. Consequently neurons are unable to maintain the pool of mature and functional lysosomes required for the autophagic clearance of α-synuclein, leading to the accumulation and spread of pathogenic α-synuclein species in the brain. Since GCase deficiency and lysosomal dysfunction occur with ageing and sporadic PD pathology, the decrease in lysosomal reformation may be a common feature in PD.
authors.affiliation: Department of Clinical Neurosciences, Institute of Neurology, University College London, Rowland Hill Street, London, UK.
authors.forename: Joana
authors.initials: J
authors.lastname: Magalhaes
authors.affiliation: Department of Clinical Neurosciences, Institute of Neurology, University College London, Rowland Hill Street, London, UK.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Gegg
authors.affiliation: Department of Clinical Neurosciences, Institute of Neurology, University College London, Rowland Hill Street, London, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Migdalska-Richards
authors.affiliation: Lipidomics Research Facility, University of the Highlands and Islands, Inverness, UK.
authors.forename: Mary K
authors.initials: MK
authors.lastname: Doherty
authors.affiliation: Lipidomics Research Facility, University of the Highlands and Islands, Inverness, UK.
authors.forename: Phillip D
authors.initials: PD
authors.lastname: Whitfield
authors.affiliation: Department of Clinical Neurosciences, Institute of Neurology, University College London, Rowland Hill Street, London, UK a.schapira@ucl.ac.uk.
authors.forename: Anthony H V
authors.initials: AH
authors.lastname: Schapira
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics; D001921:Brain / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D005776:Gaucher Disease / Q000235:genetics / Q000473:pathology; D005962:Glucosylceramidase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D010300:Parkinson Disease / Q000235:genetics* / Q000473:pathology; D051844:alpha-Synuclein / Q000235:genetics*; D020691:rab GTP-Binding Proteins / Q000235:genetics*; D000090802:rab7 GTP-Binding Proteins
chemical_list: C497604:SNCA protein, human; D051844:alpha-Synuclein; D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; C000716465:rab7 GTP-binding proteins, mouse; D005962:Glucosylceramidase; D020691:rab GTP-Binding Proteins