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VCP suppresses proteopathic seeding in neurons

Zhu, Jiang and Pittman, Sara and Dhavale, Dhruva and French, Rachel and Patterson, Jessica N. and Kaleelurrrahuman, Mohamed Salman and Sun, Yuanzi and Vaquer-Alicea, Jaime and Maggiore, Gianna and Clemen, Christoph S. and Buscher, William J. and Bieschke, Jan and Kotzbauer, Paul and Ayala, Yuna and Diamond, Marc I. and Davis, Albert A. and Weihl, Conrad (2022) VCP suppresses proteopathic seeding in neurons. Molecular Neurodegeneration, 17 (1), p. 30. Springer Nature. doi: 10.1186/s13024-022-00532-0. ISSN 1750-1326.

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Official URL: https://molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-022-00532-0

Abstract

Background: Neuronal uptake and subsequent spread of proteopathic seeds, such as alphaS (alpha-synuclein), Tau, and TDP-43, contribute to neurodegeneration. The cellular machinery participating in this process is poorly understood. One proteinopathy called multisystem proteinopathy (MSP) is associated with dominant mutations in Valosin Containing Protein (VCP). MSP patients have muscle and neuronal degeneration characterized by aggregate pathology that can include alphaS, Tau and TDP-43. Methods: We performed a fluorescent cell sorting based genome-wide CRISPR-Cas9 screen in alphaS biosensors. alphaS and TDP-43 seeding activity under varied conditions was assessed using FRET/Flow biosensor cells or immuno fluorescence for phosphorylated alphaS or TDP-43 in primary cultured neurons. We analyzed in vivo seeding activity by immunostaining for phosphorylated alphaS following intrastriatal injection of alphaS seeds in control or VCP disease mutation carrying mice. Results: One hundred fifty-four genes were identified as suppressors of alphaS seeding. One suppressor, VCP when chemically or genetically inhibited increased alphaS seeding in cells and neurons. This was not due to an increase in alphaS uptake or alphaS protein levels. MSP-VCP mutation expression increased alphaS seeding in cells and neurons. Intrastriatal injection of alphaS preformed fibrils (PFF) into VCP-MSP mutation carrying mice increased phospho alphaS expression as compared to control mice. Cells stably expressing fluorescently tagged TDP-43 C-terminal fragment FRET pairs (TDP-43 biosensors) generate FRET when seeded with TDP-43 PFF but not monomeric TDP-43. VCP inhibition or MSP-VCP mutant expression increases TDP-43 seeding in TDP-43 biosensors. Similarly, treatment of neurons with TDP-43 PFFs generates high molecular weight insoluble phosphorylated TDP-43 after 5days. This TDP-43 seed dependent increase in phosphorylated TDP-43 is further augmented in MSP-VCP mutant expressing neurons. Conclusion: Using an unbiased screen, we identified the multifunctional AAA ATPase VCP as a suppressor of alphaS and TDP-43 aggregate seeding in cells and neurons. VCP facilitates the clearance of damaged lysosomes via lysophagy. We propose that VCPs surveillance of permeabilized endosomes may protect against the proteopathic spread of pathogenic protein aggregates. The spread of distinct aggregate species may dictate the pleiotropic phenotypes and pathologies in VCP associated MSP.

Item URL in elib:https://elib.dlr.de/190170/
Document Type:Article
Title:VCP suppresses proteopathic seeding in neurons
Authors:
AuthorsInstitution or Email of AuthorsAuthor's ORCID iDORCID Put Code
Zhu, JiangWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Pittman, SaraWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Dhavale, DhruvaWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
French, RachelSaint Louis University School of Medicine, Edward A. Doisy Department of Biochemistry and Molecular Biology, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Patterson, Jessica N.Washington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Kaleelurrrahuman, Mohamed SalmanWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Sun, YuanziUniversity College London, UCL Institute of Prion Diseases, Medical Research Council Prion Unit, London, UKUNSPECIFIEDUNSPECIFIED
Vaquer-Alicea, JaimeUniversity of Texas Southwestern Medical Center, Peter O Donnell Jr. Brain Institute, Center for Alzheimers and Neurodegenerative Diseases, Dallas, TX, USAUNSPECIFIEDUNSPECIFIED
Maggiore, GiannaUniversity of Texas Southwestern Medical Center, Peter O Donnell Jr. Brain Institute, Center for Alzheimers and Neurodegenerative Diseases, Dallas, TX, USAUNSPECIFIEDUNSPECIFIED
Clemen, Christoph S.German Aerospace Center (DLR), Institute of Aerospace Medicine, 51147 Cologne, Germanyhttps://orcid.org/0000-0002-1291-4219UNSPECIFIED
Buscher, William J.Washington University School of Medicine, Department of Genetics, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Bieschke, JanUniversity College London, UCL Institute of Prion Diseases, Medical Research Council Prion Unit, London, UKUNSPECIFIEDUNSPECIFIED
Kotzbauer, PaulWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Ayala, YunaSaint Louis University School of Medicine, Edward A. Doisy Department of Biochemistry and Molecular Biology, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Diamond, Marc I.University of Texas Southwestern Medical Center, Peter O Donnell Jr. Brain Institute, Center for Alzheimers and Neurodegenerative Diseases, Dallas, TX, USAUNSPECIFIEDUNSPECIFIED
Davis, Albert A.Washington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Weihl, ConradWashington University School of Medicine, Department of Neurology, Hope Center for Neurological Diseases, St Louis, MO, USAUNSPECIFIEDUNSPECIFIED
Date:12 April 2022
Journal or Publication Title:Molecular Neurodegeneration
Refereed publication:Yes
Open Access:Yes
Gold Open Access:Yes
In SCOPUS:Yes
In ISI Web of Science:Yes
Volume:17
DOI:10.1186/s13024-022-00532-0
Page Range:p. 30
Publisher:Springer Nature
ISSN:1750-1326
Status:Published
Keywords:CRISPR screen, Seeding, Alpha-synuclein, TDP-43, Frontotemporal dementia, VCP, Valosin, neurons
HGF - Research field:Aeronautics, Space and Transport
HGF - Program:Space
HGF - Program Themes:Research under Space Conditions
DLR - Research area:Raumfahrt
DLR - Program:R FR - Research under Space Conditions
DLR - Research theme (Project):R - Muscle Mechanics and Metabolism
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Gravitational Biology
Deposited By: Chiodo, Annette
Deposited On:17 Nov 2022 09:36
Last Modified:25 Nov 2022 13:07

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