Cellular Prion Protein Mediates α-Synuclein Uptake, Localization, and Toxicity In Vitro and In Vivo

Tobias Thom; Matthias Schmitz; Anna-Lisa Fischer; Angela Correia; Susana Correia; Franc Llorens; Anna-Villar Pique; Wiebke Möbius; Renato Domingues; Saima Zafar; Erik Stoops; Christopher J Silva; Andre Fischer; Tiago F Outeiro; Inga Zerr

doi:10.1002/mds.28774

Cellular Prion Protein Mediates α-Synuclein Uptake, Localization, and Toxicity In Vitro and In Vivo

Mov Disord. 2022 Jan;37(1):39-51. doi: 10.1002/mds.28774. Epub 2021 Aug 27.

Authors

Tobias Thom¹, Matthias Schmitz¹, Anna-Lisa Fischer¹, Angela Correia¹, Susana Correia¹, Franc Llorens^{1

2

3}, Anna-Villar Pique^{1

2

3}, Wiebke Möbius⁴, Renato Domingues⁵, Saima Zafar^{1

6}, Erik Stoops⁷, Christopher J Silva⁸, Andre Fischer^{9

10

11}, Tiago F Outeiro^{5

12

13}, Inga Zerr¹

Affiliations

¹ Department of Neurology, University Medical Center Göttingen and the German Center for Neurodegenerative Diseases, Göttingen, Germany.
² Network Center for Biomedical Research of Neurodegenerative Diseases (CIBERNED), Institute Carlos III, Madrid, Spain.
³ Bellvitge Biomedical Research Institute, L'Hospitalet de Llobregat, Spain.
⁴ Department for Neurogenetics, EM Core Unit Max Planck Institute for Experimental Medicine, Göttingen, Germany.
⁵ Department of Experimental Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany.
⁶ Biomedical Engineering and Sciences Department, School of Mechanical and Manufacturing Engineering, National University of Sciences and Technology, Islamabad, Pakistan.
⁷ ADx NeuroSciences, Ghent, Belgium.
⁸ Produce Safety & Microbiology Research Unit, Western Regional Research Center, United States Department of Agriculture, Agricultural Research Service, Albany, California, USA.
⁹ Department for Epigenetics and Systems Medicine in Neurodegenerative Diseases, German Center for Neurodegenerative Diseases, Göttingen, Germany.
¹⁰ Department of Psychiatry and Psychotherapy, University Medical Center Göttingen, Göttingen, Germany.
¹¹ Cluster of Excellence "Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells" (MBExC), University of Göttingen, Göttingen, Germany.
¹² Max Planck Institute for Experimental Medicine, Goettingen, Germany.
¹³ Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.

PMID: 34448510
DOI: 10.1002/mds.28774

Abstract

Background: The cellular prion protein (PrP^C ) is a membrane-bound, multifunctional protein mainly expressed in neuronal tissues. Recent studies indicate that the native trafficking of PrP^C can be misused to internalize misfolded amyloid beta and α-synuclein (aSyn) oligomers.

Objectives: We define PrP^C 's role in internalizing misfolded aSyn in α-synucleinopathies and identify further involved proteins.

Methods: We performed comprehensive behavioral studies on four transgenic mouse models (ThySyn and ThySynPrP00, TgM83 and TgMPrP00) at different ages. We developed PrP^C -(over)-expressing cell models (cell line and primary cortical neurons), used confocal laser microscopy to perform colocalization studies, applied mass spectrometry to identify interactomes, and determined disassociation constants using surface plasmon resonance (SPR) spectroscopy.

Results: Behavioral deficits (memory, anxiety, locomotion, etc.), reduced lifespans, and higher oligomeric aSyn levels were observed in PrP^C -expressing mice (ThySyn and TgM83), but not in homologous Prnp ablated mice (ThySynPrP00 and TgMPrP00). PrP^C colocalized with and facilitated aSyn (oligomeric and monomeric) internalization in our cell-based models. Glimepiride treatment of PrP^C -overexpressing cells reduced aSyn internalization in a dose-dependent manner. SPR analysis showed that the binding affinity of PrP^C to monomeric aSyn was lower than to oligomeric aSyn. Mass spectrometry-based proteomic studies identified clathrin in the immunoprecipitates of PrP^C and aSyn. SPR was used to show that clathrin binds to recombinant PrP, but not aSyn. Experimental disruption of clathrin-coated vesicles significantly decreased aSyn internalization.

Conclusion: PrP^C 's native trafficking can be misused to internalize misfolded aSyn through a clathrin-based mechanism, which may facilitate the spreading of pathological aSyn. Disruption of aSyn-PrP^C binding is, therefore, an appealing therapeutic target in α-synucleinopathies. © 2021 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Keywords: cellular prion protein; α-synuclein; α-synucleinopathies.

Publication types

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

MeSH terms

Amyloid beta-Peptides
Animals
Mice
Prion Proteins
Proteomics
Synucleinopathies*
alpha-Synuclein* / metabolism

Substances

Amyloid beta-Peptides
Prion Proteins
alpha-Synuclein