eprintid: 10146610 rev_number: 12 eprint_status: archive userid: 699 dir: disk0/10/14/66/10 datestamp: 2022-04-11 12:48:17 lastmod: 2022-04-11 12:55:42 status_changed: 2022-04-11 12:55:42 type: article metadata_visibility: show sword_depositor: 699 creators_name: Galvagnion, Céline creators_name: Marlet, Frederik Ravnkilde creators_name: Cerri, Silvia creators_name: Schapira, Anthony HV creators_name: Blandini, Fabio creators_name: Di Monte, Donato A title: Sphingolipid changes in Parkinson L444P GBA mutation fibroblasts promote α-synuclein aggregation ispublished: inpress divisions: C07 divisions: F84 divisions: B02 divisions: UCL divisions: D07 keywords: GBA , Parkinson’s disease, fibroblasts, lipidomics, α-synuclein note: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/ by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com abstract: Intraneuronal accumulation of aggregated α-synuclein is a pathological hallmark of Parkinson's disease. Therefore, mechanisms capable of promoting α-synuclein deposition bear important pathogenetic implications. Mutations of the glucocerebrosidase 1 (GBA) gene represent a prevalent Parkinson's disease risk factor. They are associated with loss of activity of a key enzyme involved in lipid metabolism, glucocerebrosidase, supporting a mechanistic relationship between abnormal α-synuclein-lipid interactions and the development of Parkinson pathology. In this study, the lipid membrane composition of fibroblasts isolated from control subjects, patients with idiopathic Parkinson's disease and Parkinson's disease patients carrying the L444P GBA mutation (PD-GBA) was assayed using shotgun lipidomics. The lipid profile of PD-GBA fibroblasts differed significantly from that of control and idiopathic Parkinson's disease cells. It was characterized by an overall increase in sphingolipid levels. It also featured a significant increase in the proportion of ceramide, sphingomyelin and hexosylceramide molecules with shorter chain length and a decrease in the percentage of longer-chain sphingolipids. The extent of this shift was correlated to the degree of reduction of fibroblast glucocerebrosidase activity. Lipid extracts from control and PD-GBA fibroblasts were added to recombinant α-synuclein solutions. The kinetics of α-synuclein aggregation were significantly accelerated after addition of PD-GBA extracts as compared to control samples. Amyloid fibrils collected at the end of these incubations contained lipids, indicating α-synuclein-lipid co-assembly. Lipids extracted from α-synuclein fibrils were also analysed by shotgun lipidomics. Data revealed that the lipid content of these fibrils was significantly enriched by shorter-chain sphingolipids. In a final set of experiments, control and PD-GBA fibroblasts were incubated in the presence of the small molecule chaperone ambroxol. This treatment restored glucocerebrosidase activity and sphingolipid levels and composition of PD-GBA cells. It also reversed the pro-aggregation effect that lipid extracts from PD-GBA fibroblasts had on α-synuclein. Taken together, the findings of this study indicate that the L444P GBA mutation and consequent enzymatic loss are associated with a distinctly altered membrane lipid profile that provides a biological fingerprint of this mutation in Parkinson fibroblasts. This altered lipid profile could also be an indicator of increased risk for α-synuclein aggregate pathology. date: 2022-04-01 date_type: published publisher: Oxford University Press (OUP) official_url: https://doi.org/10.1093/brain/awab371 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1947735 doi: 10.1093/brain/awab371 medium: Print-Electronic pii: 6555865 lyricists_name: Schapira, Anthony lyricists_id: AHVSC78 actors_name: Flynn, Bernadette actors_id: BFFLY94 actors_role: owner funding_acknowledgements: H2020-MSCA, 706551 [Marie Skłodowska-Curie Actions-Individual Fellowship]; R314-2018-3493 [Lundbeck Foundation]; CF19-0382 [Carlsberg Foundation]; IMI 821522, PD-MitoQUANT [EU Innovative Medicines Initiative]; JPND 01ED2005B, GBA-PaCTS [EU Joint Programme-Neurodegenerative Disease Research]; ASAP-000420 [The Michael J. Fox Foundation for Parkinson's Research and the Aligning Science Across Parkinson's] full_text_status: public publication: Brain event_location: England citation: Galvagnion, Céline; Marlet, Frederik Ravnkilde; Cerri, Silvia; Schapira, Anthony HV; Blandini, Fabio; Di Monte, Donato A; (2022) Sphingolipid changes in Parkinson L444P GBA mutation fibroblasts promote α-synuclein aggregation. Brain 10.1093/brain/awab371 <https://doi.org/10.1093/brain%2Fawab371>. (In press). Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10146610/1/awab371.pdf