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shRNA-Based Screen Identifies Endocytic Recycling Pathway Components That Act as Genetic Modifiers of Alpha-Synuclein Aggregation, Secretion and Toxicity

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posted on 10.05.2016, 11:24 by Susana A. Gonçalves, Diana Macedo, Helena Raquel, Pedro D. Simões, Flaviano Giorgini, José S. Ramalho, Duarte C. Barral, Luís Ferreira Moita, Tiago Fleming Outeiro
Alpha-Synuclein (aSyn) misfolding and aggregation is common in several neurodegenerative diseases, including Parkinson's disease and dementia with Lewy bodies, which are known as synucleinopathies. Accumulating evidence suggests that secretion and cell-to-cell trafficking of pathological forms of aSyn may explain the typical patterns of disease progression. However, the molecular mechanisms controlling aSyn aggregation and spreading of pathology are still elusive. In order to obtain unbiased information about the molecular regulators of aSyn oligomerization, we performed a microscopy-based large-scale RNAi screen in living cells. Interestingly, we identified nine Rab GTPase and kinase genes that modulated aSyn aggregation, toxicity and levels. From those, Rab8b, Rab11a, Rab13 and Slp5 were able to promote the clearance of aSyn inclusions and rescue aSyn induced toxicity. Furthermore, we found that endocytic recycling and secretion of aSyn was enhanced upon Rab11a and Rab13 expression in cells accumulating aSyn inclusions. Overall, our study resulted in the identification of new molecular players involved in the aggregation, toxicity, and secretion of aSyn, opening novel avenues for our understanding of the molecular basis of synucleinopathies.

Funding

SAG was supported by Axa Research Fund (www.axa-research.org)) and Fundação para a Ciência e Tecnologia (www.fct.pt), SFRH/BD/79337/2011. DM was supported by and Fundação para a Ciência e Tecnologia (www.fct.pt), SFRH/BD/73429/2010. DCB was supported by a Marie Curie International Reintegration Grant (http://cordis.europa.eu/mariecurie-actions/irg/home.html), PIRG05-GA-2009-247726. JSR was supported by Fundação para a Ciência e Tecnologia (www.fct.pt/), PTDC_SAU_OSM_104668/2008. Work in LFM´s laboratory related to this project was supported by Fundação Luso-Americana para o Desenvolvimento (FLAD,www.flad.pt/en). LFM is an FCT Investigator and receives financial support from the European Research Council (ERC-2014-CoG 647888-iPROTECTION). This work was supported in part by funding from Parkinson’s UK to FG and TFO. TFO was also supported by a grant from the Michael J. Fox Foundation (www.michaeljfox.org) and is currently supported by the DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (www.neurodegeneration.uni-goettingen.de).

History

Citation

PLoS Genetics, 2016, 12 (4), pp. e1005995

Author affiliation

/Organisation/COLLEGE OF MEDICINE, BIOLOGICAL SCIENCES AND PSYCHOLOGY/MBSP Non-Medical Departments/Department of Genetics

Version

VoR (Version of Record)

Published in

PLoS Genetics

Publisher

Public Library of Science

issn

1553-7390

eissn

1553-7404

Acceptance date

28/03/2016

Copyright date

2016

Available date

10/05/2016

Publisher version

http://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1005995

Language

en

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