Alpha-Synuclein Cell-to-Cell Transfer and Seeding in Grafted Dopaminergic Neurons In Vivo

Alpha-synuclein (α-syn) and leucine-rich repeat kinase 2 (LRRK2) play crucial roles in Parkinson’s disease (PD). They may functionally interact to induce the degeneration of dopaminergic (DA) neurons via mechanisms that are not yet fully understood. We previously showed that the C-terminal portion of LRRK2 (ΔLRRK2) with the G2019S mutation (ΔLRRK2G2019S) was sufficient to induce neurodegeneration of DA neurons in vivo, suggesting that mutated LRRK2 induces neurotoxicity through mechanisms that are (i) independent of the N-terminal domains and (ii) “cell-autonomous”. Here, we explored whether ΔLRRK2G2019S could modify α-syn toxicity through these two mechanisms. We used a co-transduction approach in rats with AAV vectors encoding ΔLRRK2G2019S or its “dead” kinase form, ΔLRRK2DK, and human α-syn with the A53T mutation (AAV-α-synA53T). Behavioral and histological evaluations were performed at 6- and 15-weeks post-injection. Results showed that neither form of ΔLRRK2 alone induced the degeneration of neurons at these post-injection time points. By contrast, injection of AAV-α-synA53T alone resulted in motor signs and degeneration of DA neurons. Co-injection of AAV-α-synA53T with AAV-ΔLRRK2G2019S induced DA neuron degeneration that was significantly higher than that induced by AAV-α-synA53T alone or with AAV-ΔLRRK2DK. Thus, mutated α-syn neurotoxicity can be enhanced by the C-terminal domain of LRRK2G2019 alone, through cell-autonomous mechanisms.

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Aplp1 and the Aplp1-Lag3 Complex facilitates transmission of pathologic alpha-synuclein

10.1101/2021.05.01.442157 ◽

2021 ◽

Author(s):

Xiaobo Mao ◽

Hao Gu ◽

Donghoon Kim ◽

Yasuyoshi Kimura ◽

Ning Wang ◽

...

Keyword(s):

Parkinson Disease ◽

Molecular Mechanism ◽

Amyloid Beta ◽

Dopaminergic Neurons ◽

Lymphocyte Activation ◽

Alpha Synuclein ◽

Therapeutic Strategies ◽

Behavioral Deficits ◽

Lymphocyte Activation Gene

Pathologic alpha-synuclein (alpha-syn) spreads from cell-to-cell, in part, through binding to the lymphocyte-activation gene 3 (Lag3). Here we report that amyloid beta precursor-like protein 1 (Aplp1) forms a complex with Lag3 that facilitates the binding, internalization, transmission, and toxicity of pathologic alpha-syn. Deletion of both Aplp1 and Lag3 eliminates the loss of dopaminergic neurons and the accompanying behavioral deficits induced by alpha-syn preformed fibrils (PFF). Anti-Lag3 prevents the internalization of alpha-syn PFF by disrupting the interaction of Aplp1 and Lag3, and blocks the neurodegeneration induced by alpha-syn PFF in vivo. The identification of Aplp1 and the interplay with Lag3 for alpha-syn PFF induced pathology advances our understanding of the molecular mechanism of cell-to-cell transmission of pathologic alpha-syn and provides additional targets for therapeutic strategies aimed at preventing neurodegeneration in Parkinson disease and related alpha-synucleinopathies.

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RIT2 reduces LRRK2 kinase activity and protects against alpha-synuclein neuropathology

10.1101/2020.10.21.348144 ◽

2020 ◽

Author(s):

Julia Obergasteiger ◽

Anne-Marie Castonguay ◽

Giulia Frapporti ◽

Evy Lobbestael ◽

Veerle Baekelandt ◽

...

Keyword(s):

Risk Factor ◽

Substantia Nigra ◽

Dopaminergic Neurons ◽

Kinase Activity ◽

Alpha Synuclein ◽

The Novel ◽

Lrrk2 Kinase ◽

Novel Strategy ◽

Lrrk2 Kinase Activity

AbstractIn Parkinson’s disease (PD) misfolded alpha-synuclein (aSyn) accumulates in the substantia nigra, where dopaminergic neurons are progressively lost. The mechanisms underlying aSyn pathology are still unclear but hypothesized to involve the autophagy lysosome pathways (ALP). LRRK2 mutations are a major cause of familial and sporadic PD, hyperactivate kinase activity and its pharmacological inhibition reduces pS129-aSyn inclusions. We observed selective downregulation of the novel PD risk factor RIT2 in G2019S-LRRK2 expressing cells. Here we studied whether RIT2 could modulate LRRK2 kinase activity. RIT2 overexpression in G2019S-LRRK2 cells rescued ALP abnormalities and diminished aSyn inclusions. In vivo, viral mediated overexpression of RIT2 operated neuroprotection against AAV-A53T-aSyn. Furthermore, RIT2 overexpression prevented the A53T-aSyn-dependent increase of LRRK2 kinase activity in vivo. Our data indicate that RIT2 inhibits overactive LRRK2 to ameliorate ALP impairment and counteract aSyn aggregation and related deficits. Targeting RIT2 could represent a novel strategy to combat neuropathology in familial and idiopathic PD.

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A new model to study cell-to-cell transfer of αSynuclein in vivo

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2018.07.053 ◽

2018 ◽

Vol 503 (3) ◽

pp. 1385-1393 ◽

Cited By ~ 2

Author(s):

Gabriela Mercado ◽

Nélida López ◽

Alexis Martínez ◽

Sergio P. Sardi ◽

Claudio Hetz

Keyword(s):

Cell Transfer ◽

New Model ◽

Cell To Cell Transfer

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Mitochondrial alpha-synuclein accumulation impairs complex I function in dopaminergic neurons and results in increased mitophagy in vivo

Neuroscience Letters ◽

10.1016/j.neulet.2010.09.061 ◽

2010 ◽

Vol 486 (3) ◽

pp. 235-239 ◽

Cited By ~ 232

Author(s):

Shankar J. Chinta ◽

Jyothi K. Mallajosyula ◽

Anand Rane ◽

Julie K. Andersen

Keyword(s):

Dopaminergic Neurons ◽

Complex I ◽

Alpha Synuclein

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The Parkinson's Disease-Related Protein DJ-1 Protects Dopaminergic Neurons in vivo and Cultured Cells from Alpha-Synuclein and 6-Hydroxydopamine Toxicity

Neurodegenerative Diseases ◽

10.1159/000367993 ◽

2014 ◽

Vol 15 (1) ◽

pp. 13-23 ◽

Cited By ~ 16

Author(s):

Sara Batelli ◽

Roberto William Invernizzi ◽

Alessandro Negro ◽

Eleonora Calcagno ◽

Serena Rodilossi ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Dopaminergic Neurons ◽

Cultured Cells ◽

Alpha Synuclein ◽

Related Protein ◽

6 Hydroxydopamine

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Faculty Opinions recommendation of In vivo demonstration that alpha-synuclein oligomers are toxic.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.9204956.9798058 ◽

2011 ◽

Author(s):

Harry Ischiropoulos

Keyword(s):

Alpha Synuclein

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An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of α-synuclein

Cell Reports ◽

10.1016/j.celrep.2021.109189 ◽

2021 ◽

Vol 35 (10) ◽

pp. 109189

Author(s):

Eleanna Kara ◽

Alessandro Crimi ◽

Anne Wiedmer ◽

Marc Emmenegger ◽

Claudia Manzoni ◽

...

Keyword(s):

Cell Transfer ◽

Genomic Approach ◽

Genetic Landscape ◽

Cell To Cell Transfer

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Metabolic radiolabeling and in vivo PET imaging of cytotoxic T lymphocytes to guide combination adoptive cell transfer cancer therapy

Journal of Nanobiotechnology ◽

10.1186/s12951-021-00924-2 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Dehua Lu ◽

Yanpu Wang ◽

Ting Zhang ◽

Feng Wang ◽

Kui Li ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Solid Tumors ◽

Cytotoxic T Lymphocytes ◽

Pet Imaging ◽

Stage Migration ◽

Cell Transfer ◽

Tumor Infiltration ◽

Antitumor Effects

Abstract Background Adoptive T cell transfer-based immunotherapy yields unsatisfactory results in the treatment of solid tumors, partially owing to limited tumor infiltration and the immunosuppressive microenvironment in solid tumors. Therefore, strategies for the noninvasive tracking of adoptive T cells are critical for monitoring tumor infiltration and for guiding the development of novel combination therapies. Methods We developed a radiolabeling method for cytotoxic T lymphocytes (CTLs) that comprises metabolically labeling the cell surface glycans with azidosugars and then covalently conjugating them with 64Cu-1,4,7-triazacyclononanetriacetic acid-dibenzo-cyclooctyne (64Cu-NOTA-DBCO) using bioorthogonal chemistry. 64Cu-labeled control-CTLs and ovalbumin-specific CTLs (OVA-CTLs) were tracked using positron emission tomography (PET) in B16-OVA tumor-bearing mice. We also investigated the effects of focal adhesion kinase (FAK) inhibition on the antitumor efficacy of OVA-CTLs using a poly(lactic-co-glycolic) acid (PLGA)-encapsulated nanodrug (PLGA-FAKi). Results CTLs can be stably radiolabeled with 64Cu with a minimal effect on cell viability. PET imaging of 64Cu-OVA-CTLs enables noninvasive mapping of their in vivo behavior. Moreover, 64Cu-OVA-CTLs PET imaging revealed that PLGA-FAKi induced a significant increase in OVA-CTL infiltration into tumors, suggesting the potential for a combined therapy comprising OVA-CTLs and PLGA-FAKi. Further combination therapy studies confirmed that the PLGA-FAKi nanodrug markedly improved the antitumor effects of adoptive OVA-CTLs transfer by multiple mechanisms. Conclusion These findings demonstrated that metabolic radiolabeling followed by PET imaging can be used to sensitively profile the early-stage migration and tumor-targeting efficiency of adoptive T cells in vivo. This strategy presents opportunities for predicting the efficacy of cell-based adoptive therapies and for guiding combination regimens. Graphic Abstract

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Multiple system atrophy-associated oligodendroglial protein p25α stimulates formation of novel α-synuclein strain with enhanced neurodegenerative potential

Acta Neuropathologica ◽

10.1007/s00401-021-02316-0 ◽

2021 ◽

Author(s):

Nelson Ferreira ◽

Hjalte Gram ◽

Zachary A. Sorrentino ◽

Emil Gregersen ◽

Sissel Ida Schmidt ◽

...

Keyword(s):

Multiple System Atrophy ◽

Protein Misfolding ◽

Resonance Energy Transfer ◽

Lewy Bodies ◽

Resonance Energy ◽

Alpha Synuclein ◽

Striatal Neurons ◽

End Stage ◽

Intracellular Aggregates

AbstractPathology consisting of intracellular aggregates of alpha-Synuclein (α-Syn) spread through the nervous system in a variety of neurodegenerative disorders including Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. The discovery of structurally distinct α-Syn polymorphs, so-called strains, supports a hypothesis where strain-specific structures are templated into aggregates formed by native α-Syn. These distinct strains are hypothesised to dictate the spreading of pathology in the tissue and the cellular impact of the aggregates, thereby contributing to the variety of clinical phenotypes. Here, we present evidence of a novel α-Syn strain induced by the multiple system atrophy-associated oligodendroglial protein p25α. Using an array of biophysical, biochemical, cellular, and in vivo analyses, we demonstrate that compared to α-Syn alone, a substoichiometric concentration of p25α redirects α-Syn aggregation into a unique α-Syn/p25α strain with a different structure and enhanced in vivo prodegenerative properties. The α-Syn/p25α strain induced larger inclusions in human dopaminergic neurons. In vivo, intramuscular injection of preformed fibrils (PFF) of the α-Syn/p25α strain compared to α-Syn PFF resulted in a shortened life span and a distinct anatomical distribution of inclusion pathology in the brain of a human A53T transgenic (line M83) mouse. Investigation of α-Syn aggregates in brain stem extracts of end-stage mice demonstrated that the more aggressive phenotype of the α-Syn/p25α strain was associated with an increased load of α-Syn aggregates based on a Förster resonance energy transfer immunoassay and a reduced α-Syn aggregate seeding activity based on a protein misfolding cyclic amplification assay. When injected unilaterally into the striata of wild-type mice, the α-Syn/p25α strain resulted in a more-pronounced motoric phenotype than α-Syn PFF and exhibited a “tropism” for nigro-striatal neurons compared to α-Syn PFF. Overall, our data support a hypothesis whereby oligodendroglial p25α is responsible for generating a highly prodegenerative α-Syn strain in multiple system atrophy.

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