Mitochondrial CISD1/Cisd accumulation blocks mitophagy and genetic or pharmacological inhibition rescues neurodegenerative phenotypes in Pink1/parkin models.

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Authors
Sanchez-Martinez, Alvaro 
Pickering, Jake T 
Twyning, Madeleine J 
Terriente-Felix, Ana 
Abstract

BACKGROUND: Mitochondrial dysfunction and toxic protein aggregates have been shown to be key features in the pathogenesis of neurodegenerative diseases, such as Parkinson's disease (PD). Functional analysis of genes linked to PD have revealed that the E3 ligase Parkin and the mitochondrial kinase PINK1 are important factors for mitochondrial quality control. PINK1 phosphorylates and activates Parkin, which in turn ubiquitinates mitochondrial proteins priming them and the mitochondrion itself for degradation. However, it is unclear whether dysregulated mitochondrial degradation or the toxic build-up of certain Parkin ubiquitin substrates is the driving pathophysiological mechanism leading to PD. The iron-sulphur cluster containing proteins CISD1 and CISD2 have been identified as major targets of Parkin in various proteomic studies. METHODS: We employed in vivo Drosophila and human cell culture models to study the role of CISD proteins in cell and tissue viability as well as aged-related neurodegeneration, specifically analysing aspects of mitophagy and autophagy using orthogonal assays. RESULTS: We show that the Drosophila homolog Cisd accumulates in Pink1 and parkin mutant flies, as well as during ageing. We observed that build-up of Cisd is particularly toxic in neurons, resulting in mitochondrial defects and Ser65-phospho-Ubiquitin accumulation. Age-related increase of Cisd blocks mitophagy and impairs autophagy flux. Importantly, reduction of Cisd levels upregulates mitophagy in vitro and in vivo, and ameliorates pathological phenotypes in locomotion, lifespan and neurodegeneration in Pink1/parkin mutant flies. In addition, we show that pharmacological inhibition of CISD1/2 by rosiglitazone and NL-1 induces mitophagy in human cells and ameliorates the defective phenotypes of Pink1/parkin mutants. CONCLUSION: Altogether, our studies indicate that Cisd accumulation during ageing and in Pink1/parkin mutants is a key driver of pathology by blocking mitophagy, and genetically and pharmacologically inhibiting CISD proteins may offer a potential target for therapeutic intervention.

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Acknowledgements: We kindly thank Prof. Ian Ganley (MRC Protein Phosphorylation and Ubiquitylation Unit, University of Dundee) for proving us with the ARPE-19-mito-QC cells; Prof. David Walker and Prof. Jongkyeong Chung for providing fly lines; Prof. Jon Lane (University of Bristol) for generously donating us the hTERT-RPE1 and hTERT-RPE1-YFP-PARKIN cells; Jan Miljkovic, Jordan Morris, and especially Michele Frison for valuable input; and all the members of the Whitworth lab for discussions. We also thank Prof. Chung and Prof. Axel Methner for discussions and communicating results prior to publication. The Graphical Abstract was created with BioRender.com.

Keywords
Ageing, Autophagy, CISD1, CISD2, Cisd, Mitochondria, Mitophagy, Neurodegeneration, PINK1, Parkin, Parkinson’s disease, Animals, Humans, Aged, Mitophagy, Protein Kinases, Proteomics, Ubiquitin-Protein Ligases, Parkinson Disease, Mitochondrial Proteins, Drosophila, Mitochondria, Ubiquitins, Protein Serine-Threonine Kinases, Drosophila Proteins
Journal Title
Mol Neurodegener
Conference Name
Journal ISSN
1750-1326
1750-1326
Volume Title
19
Publisher
Springer Science and Business Media LLC
Sponsorship
Hezkuntza, Hizkuntza Politika Eta Kultura Saila, Eusko Jaurlaritza (POS_2022_2_0045)
National Science and Technology Council (110-2311-B-400 -002 -MY3)
MRC core funding (MC_UU_00028/6)