Universität zu Köln

Zhu, Jiang, Pittman, Sara, Dhavale, Dhruva ORCID: 0000-0002-0096-1085, French, Rachel, Patterson, Jessica N., Kaleelurrrahuman, Mohamed Salman, Sun, Yuanzi, Vaquer-Alicea, Jaime, Maggiore, Gianna, Clemen, Christoph S., Buscher, William J., Bieschke, Jan, Kotzbauer, Paul, Ayala, Yuna, Diamond, Marc I., Davis, Albert A. and Weihl, Conrad (2022). VCP suppresses proteopathic seeding in neurons. Mol. Neurodegener., 17 (1). LONDON: BMC. ISSN 1750-1326

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Abstract

Background Neuronal uptake and subsequent spread of proteopathic seeds, such as alpha S (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 alpha S, Tau and TDP-43. Methods We performed a fluorescent cell sorting based genome-wide CRISPR-Cas9 screen in alpha S biosensors. alpha S and TDP-43 seeding activity under varied conditions was assessed using FRET/Flow biosensor cells or immunofluorescence for phosphorylated alpha S or TDP-43 in primary cultured neurons. We analyzed in vivo seeding activity by immunostaining for phosphorylated alpha S following intrastriatal injection of alpha S seeds in control or VCP disease mutation carrying mice. Results One hundred fifty-four genes were identified as suppressors of alpha S seeding. One suppressor, VCP when chemically or genetically inhibited increased alpha S seeding in cells and neurons. This was not due to an increase in alpha S uptake or alpha S protein levels. MSP-VCP mutation expression increased alpha S seeding in cells and neurons. Intrastriatal injection of alpha S preformed fibrils (PFF) into VCP-MSP mutation carrying mice increased phospho alpha S 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 5 days. This TDP-43 seed dependent increase in phosphorlyated 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 alpha S and TDP-43 aggregate seeding in cells and neurons. VCP facilitates the clearance of damaged lysosomes via lysophagy. We propose that VCP's 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 Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Zhu, Jiang UNSPECIFIED UNSPECIFIED UNSPECIFIED Pittman, Sara UNSPECIFIED UNSPECIFIED UNSPECIFIED Dhavale, Dhruva UNSPECIFIED orcid.org/0000-0002-0096-1085 UNSPECIFIED French, Rachel UNSPECIFIED UNSPECIFIED UNSPECIFIED Patterson, Jessica N. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kaleelurrrahuman, Mohamed Salman UNSPECIFIED UNSPECIFIED UNSPECIFIED Sun, Yuanzi UNSPECIFIED UNSPECIFIED UNSPECIFIED Vaquer-Alicea, Jaime UNSPECIFIED UNSPECIFIED UNSPECIFIED Maggiore, Gianna UNSPECIFIED UNSPECIFIED UNSPECIFIED Clemen, Christoph S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Buscher, William J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Bieschke, Jan UNSPECIFIED UNSPECIFIED UNSPECIFIED Kotzbauer, Paul UNSPECIFIED UNSPECIFIED UNSPECIFIED Ayala, Yuna UNSPECIFIED UNSPECIFIED UNSPECIFIED Diamond, Marc I. UNSPECIFIED UNSPECIFIED UNSPECIFIED Davis, Albert A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Weihl, Conrad UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-690237
DOI:	10.1186/s13024-022-00532-0
Journal or Publication Title:	Mol. Neurodegener.
Volume:	17
Number:	1
Date:	2022
Publisher:	BMC
Place of Publication:	LONDON
ISSN:	1750-1326
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language INCLUSION-BODY MYOPATHY; ALPHA-SYNUCLEIN; FRONTOTEMPORAL DEMENTIA; MULTISYSTEM PROTEINOPATHY; PAGET-DISEASE; LEWY BODY; P97; CLEARANCE; PATHOLOGY; TDP-43 Multiple languages Neurosciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/69023