Rapid macropinocytic transfer of α-synuclein to lysosomes

Bayati, Armin; Banks, Emily; Han, Chanshuai; Luo, Wen; Reintsch, Wolfgang; Zorca, Cornelia E.; Shlaifer, Irina; Pellitero, Esther Del Cid; Vanderperre, Benoît; McBride, Heidi M.; Fon, Edward A.; Durcan, Thomas M.; McPherson, Peter S.

doi:10.1016/j.celrep.2022.111102

Cited by 29 publications

(33 citation statements)

References 117 publications

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“…A recent study in U2OS and iPSC-derived human dopaminergic neurons and astrocytes reported that a-syn PFF are internalized via macropinocytosis and trafficked to lysosomes and multivesicular bodies within minutes of exposure. These findings put in question previous reports that are in favor of asyn internalization via dynamin dependent endocytosis 30 .…”

Section: Introductioncontrasting

confidence: 58%

“…Monomeric a-syn (5 mg/ml) was allowed to spontaneously form fibrillar aggregates at 37°C in an orbital shaker for 5 days at 1,000 RPM. Quality control was assessed per batch using TEM to examine fibrillar morphology after negative staining, and Thioflavin-T assay to verify formation of amyloid fibrils, which were then sonicated by 40 cycles of 30 sec ON-OFF on a Bioruptor Pico (Denville, NJ) to yield fibrils of 50 to 100 nm long in hydrodynamic diameter as shown by dynamic light scattering 30,77 . Sonicated fibrils were aliquoted and stored at -80°C for short (which was not certified by peer review) is the author/funder.…”

Section: Pff Preparationmentioning

confidence: 99%

“…Several lines of evidence suggest that a-syn can be secreted via non-canonical vesicle-mediated exocytosis. While a small fraction of extracellular a-syn appears to be released together with exosomes 30,31 , other studies suggested an exosome independent unconventional protein secretion pathway termed as misfolding-associated protein secretion (MAPS) to specifically target misfolded proteins including a-syn and tau for secretion [32][33][34] . MAPS can be initiated when misfolded proteins are recruited by the endoplasmic reticulum (ER)-embedded deubiquitinase USP19.…”

Section: Introductionmentioning

confidence: 99%

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Genetic inactivation of the USP19 deubiquitinase regulates a-synuclein ubiquitination and inhibits accumulation of Lewy body like aggregates in mice

Schorova

Bédard

Khayachi

et al. 2022

Preprint

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The USP19 deubiquitinase is found in a locus associated with Parkinson's Disease (PD), interacts with heat shock proteins and promotes secretion of a-synuclein (a-syn) through the misfolding associated protein secretion (MAPS) pathway. Since these processes might modulate the processing of a-syn aggregates during the progression of PD, we tested the effect of USP19 knockout (KO) in mice expressing the A53T mutation of a-syn and in whom a-syn preformed fibrils (PFF) had been injected in the striatum. Compared to WT, KO brains showed decreased accumulation of phospho-synuclein (pSyn) positive aggregates. The improved pathology was associated with less activation of microglia, higher levels of synaptic marker proteins and improved performance in a tail suspension test. Exposure of primary neurons from WT and KO mice to PFF in vitro also led to decreased accumulation of pSyn aggregates. KO did not affect uptake of PFF in the cultured neurons. It also did not affect the propagation of aggregates as assessed by exposing WT or KO neurons to PFF and measuring pSyn positive aggregates in non-exposed adjacent neurons separated using a microfluidics device. We conclude that USP19 instead modulates intracellular dynamics of aggregates. Indeed, at the early time following PFF injection when the number of pSyn positive neurons were similar in WT and KO brains, the KO neurons contained less aggregates. KO brain aggregates stained more intensely with anti-ubiquitin antibodies. Immunoprecipitation of soluble proteins from primary neurons exposed to PFF with antibodies to ubiquitin or pSyn showed higher levels of ubiquitinated a-syn oligomeric species in the KO neurons. We propose that the improved pathology in USP19 KO brains may arise from decreased formation or enhanced clearance of the more ubiquitinated aggregates and/or enhanced disassembly towards more soluble oligomeric species. USP19 inhibition may represent a novel therapeutic approach that targets the intracellular dynamics of a-syn complexes.

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Section: Introductioncontrasting

confidence: 58%

Section: Pff Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genetic inactivation of the USP19 deubiquitinase regulates a-synuclein ubiquitination and inhibits accumulation of Lewy body like aggregates in mice

Schorova

Bédard

Khayachi

et al. 2022

Preprint

Self Cite

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“…This process encompasses a range of distinct pathways, macroautophagy, microautophagy, and chaperonemediated autophagy (CMA), to name the most relevant. In particular, aggregated forms of αS were reported to colocalize with both early and late endo/lysosomal markers (Lee et al, 2008;Brahic et al, 2016;Karpowicz et al, 2017;Bayati et al, 2022). Human αS overexpression was observed to impair macroautophagy in mammalian cells and in transgenic mice, where it affects Rab1a-specific secretory pathways, hampering omegasome (autophagosome precursor) formation (Winslow et al, 2010), as well as to damage the autophagic flux in aging adult neurons through the inhibition of mitophagosome maturation and autophagosome-mediated αS clearance (Figure 4; Sarkar et al, 2021).…”

Section: Lysosomal and Autophagy Impairmentmentioning

confidence: 99%

α-Synuclein oligomers and fibrils: partners in crime in synucleinopathies

Cecchi¹,

Bigi²,

Cascella³

2023

Neural Regeneration Research

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The misfolding and aggregation of α-synuclein is the general hallmark of a group of devastating neurodegenerative pathologies referred to as synucleinopathies, such as Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. In such conditions, a range of different misfolded aggregates, including oligomers, protofibrils, and fibrils, are present both in neurons and glial cells. Growing experimental evidence supports the proposition that soluble oligomeric assemblies, formed during the early phases of the aggregation process, are the major culprits of neuronal toxicity; at the same time, fibrillar conformers appear to be the most efficient at propagating among interconnected neurons, thus contributing to the spreading of α-synuclein pathology. Moreover, α-synuclein fibrils have been recently reported to release soluble and highly toxic oligomeric species, responsible for an immediate dysfunction in the recipient neurons. In this review, we discuss the current knowledge about the plethora of mechanisms of cellular dysfunction caused by α-synuclein oligomers and fibrils, both contributing to neurodegeneration in synucleinopathies.

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“…However, assuming that the plasma levels are indeed reflective of the amount secreted from neurons, there are a few alternative models that fit the data. Recent research has demonstrated that a-synuclein aggregates can be released extracellularly from the lysosome or acidified MVBs 73,74 . If a-synuclein pre-formed fibril transfer occurs primarily on the surface of exosomes following the fusion of lysosomes to multivesicular bodies as proposed in Bayati et al, 2022, the number of EVs secreted may be the limiting factor for the amount of secreted a-synuclein in aged LRRK2 G2019S mice.…”

Section: Avs Derived From Lrrk2 G2019smentioning

confidence: 99%

Proteomics analysis of autophagy cargos reveals distinct adaptations in PINK1 and LRRK2 models of Parkinson disease

Goldsmith

Ordureau

Stavoe

et al. 2022

Preprint

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Autophagy is a conserved and essential cellular degradation and recycling process that is required to maintain neuronal homeostasis. Genetic and pathological evidence suggest that autophagy is disrupted in Parkinson's disease (PD), a prevalent and progressive neurodegenerative disease, yet how changes in autophagy contribute to disease pathogenesis is unclear. To better understand the cellular impacts of disrupted autophagy on neuronal function, we used unbiased proteomics to compare the cargos degraded by basal autophagy in two different mouse models of PD. We isolated autophagic vesicles from the brains of PINK1 knock-out mice and LRRK2G2019S knock-in mice, and further compared these data to observations from young and old mice. We find evidence for the upregulation of adaptive pathways to remove proteins and organelles in PINK1-/- and LRRK2G2019S mice. In PINK1-/- mouse brain, impaired mitophagy leads to increased expression of components of the autophagic machinery as well as increased expression of selective adaptors for mitochondrial autophagy independent of PINK1/Parkin, including BCL2L13. In LRRK2G2019S mice, we find that the impairments to autophagosome trafficking and acidification lead to increased cargo secretion. We further compared these data sets to proteomic data comparing young and old mice. In aged mice, we find decreased engulfment of lysosomes and increased engulfment of α-synuclein, a key component of pathogenic Lewy bodies found in PD. Together, these findings highlight the engagement of distinct compensatory pathways to maintain homeostasis in the brain upon disruption of either stress-induced or basal autophagic pathways, and we begin to identify how age-related changes may place further stress on autophagy's ability to maintain homeostasis.

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Rapid macropinocytic transfer of α-synuclein to lysosomes

Cited by 29 publications

References 117 publications

Genetic inactivation of the USP19 deubiquitinase regulates a-synuclein ubiquitination and inhibits accumulation of Lewy body like aggregates in mice

Genetic inactivation of the USP19 deubiquitinase regulates a-synuclein ubiquitination and inhibits accumulation of Lewy body like aggregates in mice

α-Synuclein oligomers and fibrils: partners in crime in synucleinopathies

Proteomics analysis of autophagy cargos reveals distinct adaptations in PINK1 and LRRK2 models of Parkinson disease

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