shRNA-Based Screen Identifies Endocytic Recycling Pathway Components That Act as Genetic Modifiers of Alpha-Synuclein Aggregation, Secretion and Toxicity.
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.
Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Lisboa, Portugal.|CEDOC, Chronic Diseases Research Center, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisboa, Portugal.
Susana A
http://orcid.org/0000-0002-8025-2624
SA
Gonçalves
Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Lisboa, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de lisboa, Oeiras, Portugal.
Diana
D
Macedo
Instituto Gulbenkian de Ciência, Oeiras, Portugal.
Helena
H
Raquel
Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Lisboa, Portugal.
Pedro D
PD
Simões
Department of Genetics, University of Leicester, Leicester, United Kingdom.
Flaviano
F
Giorgini
CEDOC, Chronic Diseases Research Center, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisboa, Portugal.
José S
JS
Ramalho
CEDOC, Chronic Diseases Research Center, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisboa, Portugal.
Duarte C
DC
Barral
Instituto Gulbenkian de Ciência, Oeiras, Portugal.
Luís
L
Ferreira Moita
CEDOC, Chronic Diseases Research Center, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisboa, Portugal.|Department of Neurodegeneration and Restorative Research, University Medical Center Göttingen, Göttingen, Germany.|Max Planck Institute for Experimental Medicine, Göttingen, Germany.
Tiago Fleming
TF
Outeiro
D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1 / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics; D006801:Humans; D020961:Lewy Body Disease / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics; D015513:Oncogene Proteins / Q000235:genetics; D010300:Parkinson Disease / Q000235:genetics*; D066329:Protein Aggregates / Q000235:genetics*; D017510:Protein Folding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051844:alpha-Synuclein / Q000235:genetics* / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D000097603:Dyrk Kinases; D000099193:rab11 GTP-Binding Proteins
D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D015513:Oncogene Proteins; D066329:Protein Aggregates; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D034741:RNA, Small Interfering; D051844:alpha-Synuclein; D020691:rab GTP-Binding Proteins; D000097603:Dyrk Kinases; D000099193:rab11 GTP-Binding Proteins; C512283:CC2D1A protein, human; C529761:Rab8b protein, human; C457772:SYTL5 protein, human; C066780:Clk dual-specificity kinases; C517596:CAMK1 protein, human; C085485:RAB13 protein, human; C547812:Rab39B protein, human