Post-translational proteomics platform identifies neurite outgrowth impairments in Parkinson's disease GBA-N370S dopamine neurons

Helle Bogetofte; Brent J Ryan; Pia Jensen; Sissel I Schmidt; Dana L E Vergoossen; Mike B Barnkob; Lisa N Kiani; Uroosa Chughtai; Rachel Heon-Roberts; Maria Claudia Caiazza; William McGuinness; Ricardo Márquez-Gómez; Jane Vowles; Fiona S Bunn; Janine Brandes; Peter Kilfeather; Jack P Connor; Hugo J R Fernandes; Tara M Caffrey; Morten Meyer; Sally A Cowley; Martin R Larsen; Richard Wade-Martins

doi:10.1016/j.celrep.2023.112180

Post-translational proteomics platform identifies neurite outgrowth impairments in Parkinson's disease GBA-N370S dopamine neurons

Cell Rep. 2023 Mar 28;42(3):112180. doi: 10.1016/j.celrep.2023.112180. Epub 2023 Mar 3.

Authors

Helle Bogetofte¹, Brent J Ryan², Pia Jensen³, Sissel I Schmidt⁴, Dana L E Vergoossen⁵, Mike B Barnkob⁶, Lisa N Kiani⁵, Uroosa Chughtai⁵, Rachel Heon-Roberts⁷, Maria Claudia Caiazza⁷, William McGuinness⁷, Ricardo Márquez-Gómez⁷, Jane Vowles⁸, Fiona S Bunn⁵, Janine Brandes⁵, Peter Kilfeather⁷, Jack P Connor⁷, Hugo J R Fernandes⁷, Tara M Caffrey⁵, Morten Meyer⁹, Sally A Cowley⁸, Martin R Larsen³, Richard Wade-Martins¹⁰

Affiliations

¹ Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Parks Road, Oxford OX1 3QU, UK; Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark; Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark.
² Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Parks Road, Oxford OX1 3QU, UK. Electronic address: brent.ryan@dpag.ox.ac.uk.
³ Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark.
⁴ Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Parks Road, Oxford OX1 3QU, UK; Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark.
⁵ Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK.
⁶ Centre for Cellular Immunotherapy of Haematological Cancer Odense (CITCO), Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, 5000 Odense C, Denmark.
⁷ Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Parks Road, Oxford OX1 3QU, UK.
⁸ James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, UK.
⁹ Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark; Department of Neurology, Odense University Hospital, 5000 Odense C, Denmark.
¹⁰ Oxford Parkinson's Disease Centre, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Parks Road, Oxford OX1 3QU, UK. Electronic address: richard.wade-martins@dpag.ox.ac.uk.

PMID: 36870058
DOI: 10.1016/j.celrep.2023.112180

Abstract

Variants at the GBA locus, encoding glucocerebrosidase, are the strongest common genetic risk factor for Parkinson's disease (PD). To understand GBA-related disease mechanisms, we use a multi-part-enrichment proteomics and post-translational modification (PTM) workflow, identifying large numbers of dysregulated proteins and PTMs in heterozygous GBA-N370S PD patient induced pluripotent stem cell (iPSC) dopamine neurons. Alterations in glycosylation status show disturbances in the autophagy-lysosomal pathway, which concur with upstream perturbations in mammalian target of rapamycin (mTOR) activation in GBA-PD neurons. Several native and modified proteins encoded by PD-associated genes are dysregulated in GBA-PD neurons. Integrated pathway analysis reveals impaired neuritogenesis in GBA-PD neurons and identify tau as a key pathway mediator. Functional assays confirm neurite outgrowth deficits and identify impaired mitochondrial movement in GBA-PD neurons. Furthermore, pharmacological rescue of glucocerebrosidase activity in GBA-PD neurons improves the neurite outgrowth deficit. Overall, this study demonstrates the potential of PTMomics to elucidate neurodegeneration-associated pathways and potential drug targets in complex disease models.

Keywords: CP: Neuroscience; Parkinson’s; glucocerebrosidase; glycosylation; iPSC; lysosome; neuritogenesis; phosphoproteomics; post-translational modifications; proteomics; stem cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Dopaminergic Neurons / metabolism
Glucosylceramidase / genetics
Glucosylceramidase / metabolism
Humans
Mutation
Neuronal Outgrowth
Parkinson Disease* / genetics
Parkinson Disease* / metabolism
Protein Processing, Post-Translational
Proteomics

Substances

Glucosylceramidase
GBA protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding