[en] Fibrillary aggregation of α-synuclein in Lewy body inclusions and nigrostriatal dopaminergic neuron degeneration define Parkinson's disease neuropathology. Mutations in GBA1, encoding glucocerebrosidase, are the most frequent genetic risk factor for Parkinson's disease. However, the lack of reliable experimental models able to reproduce key neuropathological signatures has hampered clarification of the link between mutant glucocerebrosidase and Parkinson's disease pathology. Here, we describe an innovative protocol for the generation of human induced pluripotent stem cell-derived midbrain organoids containing dopaminergic neurons with nigral identity that reproduce characteristics of advanced maturation. When applied to patients with GBA1-related Parkinson's disease, this method enabled the differentiation of midbrain organoids recapitulating dopaminergic neuron loss and fundamental features of Lewy pathology observed in human brains, including the generation of α-synuclein fibrillary aggregates with seeding activity that also propagate pathology in healthy control organoids. Concurrently, we found that the retention of mutant glucocerebrosidase in the endoplasmic reticulum and increased levels of its substrate, glucosylceramide, are determinants of α-synuclein aggregation into Lewy body-like inclusions, and the reduction of glucocerebrosidase activity accelerated α-synuclein pathology by promoting fibrillary α-synuclein deposition. Finally, we demonstrated the efficacy of ambroxol and GZ667161 (two modulators of the glucocerebrosidase pathway in clinical development for the treatment of GBA1-related Parkinson's disease) in reducing α-synuclein pathology in this model, supporting the use of midbrain organoids as a relevant preclinical platform for investigational drug screening.
Disciplines :
Life sciences: Multidisciplinary, general & others
Author, co-author :
Frattini, Emanuele ; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
Faustini, Gaia; Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy
Lopez, Gianluca ; Division of Pathology, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Milan 20122, Italy
Carsana, Emma Veronica; Department of Medical Biotechnology and Translational Medicine, University of Milano, Milan 20054, Italy
Tosi, Mattia; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
Trezzi, Ilaria; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
Magni, Manuela; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
Soldà, Giulia ; Department of Biomedical Sciences, Humanitas University, Pieve Emanuele, Milan 20072, Italy ; Medical Genetics and RNA Biology Unit, IRCCS Humanitas Research Hospital, Rozzano, Milan 20089, Italy
Straniero, Letizia; Department of Biomedical Sciences, Humanitas University, Pieve Emanuele, Milan 20072, Italy ; Medical Genetics and RNA Biology Unit, IRCCS Humanitas Research Hospital, Rozzano, Milan 20089, Italy
Facchi, Daniele; Department of Biomedical Sciences, Humanitas University, Pieve Emanuele, Milan 20072, Italy ; Medical Genetics and RNA Biology Unit, IRCCS Humanitas Research Hospital, Rozzano, Milan 20089, Italy
Samarani, Maura; Unité de Trafic Membranaire et Pathogénèse, Département de Biologie Cellulaire et de l'Infection, Institut Pasteur, Paris 75015, France
Martá-Ariza, Mitchell; Center for Cognitive Neurology, Department of Neurology, New York University Grossman School of Medicine, New York, NY 10016, USA ; Institut de Neurociències, Universitat Autònoma de Barcelona, Barcelona 08193, Spain
De Luca, Chiara Maria Giulia; Division of Neurology 5 and Neuropathology, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan 20133, Italy
Vezzoli, Elena; Advanced Light and Electron Microscopy BioImaging Centre (ALEMBIC), IRCCS San Raffaele Scientific Institute, Milan 20132, Italy
Pittaro, Alessandra; Division of Pathology, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Milan 20122, Italy
Stepanyan, Astghik; Unità Operativa Complessa, Chirurgia Generale 3, University Hospital of Padua, Padua 35128, Italy
Silipigni, Rosamaria; Laboratory of Medical Genetics, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
ROSETY, Isabel ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine > Developmental and Cellular Biology > Team Jens Christian SCHWAMBORN
SCHWAMBORN, Jens Christian ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Developmental and Cellular Biology
Sardi, Sergio Pablo; Rare and Neurological Diseases Therapeutic Area, Sanofi, Framingham, MA 01701, USA
Moda, Fabio ; Division of Neurology 5 and Neuropathology, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan 20133, Italy
Corti, Stefania ; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy ; Department of Pathophysiology and Transplantation (DEPT), Dino Ferrari Centre, University of Milan, Milan 20122, Italy
Comi, Giacomo P; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy ; Department of Pathophysiology and Transplantation (DEPT), Dino Ferrari Centre, University of Milan, Milan 20122, Italy
Blandini, Fabio; Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
Tritsch, Nicolas X; Neuroscience Institute, New York University Grossman School of Medicine, New York, NY 10016, USA ; Fresco Institute for Parkinson's and Movement Disorders, New York University Langone Health, New York, NY 10017, USA
Bortolozzi, Mario ; Department of Physics and Astronomy 'G. Galilei', University of Padua, Padua 35131, Italy ; Veneto Institute of Molecular Medicine (VIMM), Padua 35129, Italy
Ferrero, Stefano; Division of Pathology, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Milan 20122, Italy ; Department of Biomedical, Surgical, and Dental Sciences, University of Milan, Milan 20122, Italy
Cribiù, Fulvia Milena; Division of Pathology, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Milan 20122, Italy
Wisniewski, Thomas ; Center for Cognitive Neurology, Department of Neurology, New York University Grossman School of Medicine, New York, NY 10016, USA ; Department of Pathology, New York University Grossman School of Medicine, New York, NY 10016, USA ; Department of Psychiatry, New York University Grossman School of Medicine, New York, NY 10016, USA
Asselta, Rosanna; Department of Biomedical Sciences, Humanitas University, Pieve Emanuele, Milan 20072, Italy ; Medical Genetics and RNA Biology Unit, IRCCS Humanitas Research Hospital, Rozzano, Milan 20089, Italy
Aureli, Massimo; Department of Medical Biotechnology and Translational Medicine, University of Milano, Milan 20054, Italy
Bellucci, Arianna; Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy
Di Fonzo, Alessio ; Neurology Unit, IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Milan 20122, Italy
PRIN Fondazione Cariplo Italian Ministry of Health RRC Italian Ministry of Health NIH NIA NIA Fondazione Grigioni per il Morbo di Parkinson Fresco Institute for Parkinson’s and Movement Disorders Dino Ferrari Center Mizutani Foundation for Glycoscience
Funding text :
Ministero dell\u2019Universit\u00E0 e della Ricerca 2017228L3J (R.A., M.A. and M.B.); Ministero della Salute Ricerca Corrente (A.D.F. and F.M.); Fondazione Cariplo 2015\u20131017 (M.A.); Ministero della Salute GR-2021-12372019 (F.M.); Ministero della Salute PNRRMAD-2022-12376035 (F.M.); National Institutes of Health/National Institute of Aging P30AG066512 (T.W.); National Institutes of Health/National Institute of Aging P01AG060882 (T.W.); Fondazione Grigioni per il Morbo di Parkinson (L.S.); Fondazione Regionale per la Ricerca Biomedica 825575 (A.D.F.); Mizutani Foundation for Glycoscience 220054 (M.A.).
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