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Polymorphic USP8 allele promotes Parkinson’s disease by inducing the accumulation of α-synuclein through deubiquitination

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Abstract

Parkinson’s disease (PD) is one of the most common neuro-degenerative diseases characterized by α-synuclein accumulation and degeneration of dopaminergic neurons. Employing genome-wide sequencing, we identified a polymorphic USP8 allele (USP8D442G) significantly enriched in Chinese PD patients. To test the involvement of this polymorphism in PD pathogenesis, we derived dopaminergic neurons (DAn) from human-induced pluripotent stem cells (hiPSCs) reprogrammed from fibroblasts of PD patients harboring USP8D442G allele and their healthy siblings. In addition, we knock-in D442G polymorphic site into the endogenous USP8 gene of human embryonic stem cells (hESCs) and derived DAn from these knock-in hESCs to explore their cellular phenotypes and molecular mechanism. We found that expression of USP8D442G in DAn induces the accumulation and abnormal subcellular localization of α-Synuclein (α-Syn). Mechanistically, we demonstrate that D442G polymorphism enhances the interaction between α-Syn and USP8 and thus increases the K63-specific deubiquitination and stability of α-Syn . We discover a pathogenic polymorphism for PD that represent a promising therapeutic and diagnostic target for PD.

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Availability of data and materials

The published article includes all datasets generated or analyzed during this study. The whole-genome sequencing data can be accessed from Genome Sequence Archive (https://ngdc.cncb.ac.cn/gsa, accession number: HRA002315). All datasets supporting this study are available from the lead contact, Dr. Yang Xu (xuyang2020@zju.edu.cn) upon request.

Abbreviations

α-Syn:

Alpha synuclein

USP8:

Ubiquitin specific protease-8

WT:

Wild type

MT:

USP8 D442G mutation

PD:

Parkinson’s disease

IP:

Immunoprecipitation

IB:

Immunoblotting

WB:

Western blot

DUBs:

Deubiquitinases

IF:

Immunofluorescence

TH:

Tyrosine hydroxylase

TUJ1:

Tubulin beta 3 class III

iPSC:

Induced pluripotent stem cells

NSC:

Neural stem cells

hESC:

Human embryonic stem cell

DAn:

Midbrain dopaminergic neuron cell

EV:

Empty vector

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Acknowledgements

We thank Dr. X. Luo for collecting cells from PD patients and healthy siblings, Drs. Z. Huang and J. Wang for helping the analysis of sequencing data.

Funding

This work was supported by the National Natural Science Foundation of China (Nos. 81930084, U1601222), National Key Research and Development Program of China (2022YFC3401600), the Fundamental Research Funds for the Central Universities (No. 2021FZZX001-42), Ministry of Science and Technology of China (2103ZX10002008002), the Key Research and Development Program of Guangdong Province (2019B020235003), Department of Science and Technology of Guangdong Province (2020A1515110450), Traditional Chinese Medicine Bureau Of Guangdong Province of China (20211183, 20225009), the Special Project of State Key Laboratory of Dampness Syndrome of Chinese Medicine (SZ2021KF15) and the Specific Research Fund for TCM Science and Technology of Guangdong Provincial Hospital of Chinese Medicine (YN2015QN04 and 2020KT1079).

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Authors and Affiliations

  1. State Key Laboratory of Dampness Syndrome of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China

    Shouhai Wu

  2. Center for Regenerative and Translational Medicine, Guangdong Provincial Academy of Chinese Medical Sciences, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China

    Shouhai Wu & Tongxiang Lin

  3. College of Animal Sciences, Fujian Agriculture and Forestory University, 15 ShangXiaDian Road, CangShan District, Fuzhou City, Fujian Province, China

    Tongxiang Lin

  4. Department of Cardiology, Cardiovascular Key Lab of Zhejiang Province, State Key Laboratory of Transvascular Implantation Devices, The Second Affiliated Hospital, Zhejiang University School of Medicine, Zhejiang University, Hangzhou, 310009, Zhejiang, China

    Yang Xu

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Contributions

YX conceptualized, designed, and supervised the research; SW and TL conducted the experiments.; YX, SW analyzed the data; YX and SW wrote the manuscript. All authors revised and approved the manuscript.

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Correspondence to Yang Xu.

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The study was approved by the Guangdong Provincial Hospital of Traditional Chinese Medicine (TCM) Board of Ethics.

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Wu, S., Lin, T. & Xu, Y. Polymorphic USP8 allele promotes Parkinson’s disease by inducing the accumulation of α-synuclein through deubiquitination. Cell. Mol. Life Sci. 80, 363 (2023). https://doi.org/10.1007/s00018-023-05006-0

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