Seeding the aggregation of TDP-43 requires post-fibrillization proteolytic cleavage

Nature Neuroscience, Journal Year: 2023, Volume and Issue: 26(6), P. 983 - 996

Published: May 29, 2023

Abstract Despite the strong evidence linking transactive response DNA-binding protein 43 (TDP-43) aggregation to pathogenesis of frontotemporal lobar degeneration with TDP-43, amyotrophic lateral sclerosis and several neurodegenerative diseases, our knowledge sequence structural determinants its neurotoxicity remains incomplete. Herein, we present a new method for producing recombinant full-length TDP-43 filaments that exhibit morphological features similar those brain-derived filaments. We show contain β-sheet-rich helical amyloid core is fully buried by flanking structured domains protein. demonstrate proteolytic cleavage exposure this are necessary propagating pathology enhancing seeding aggregates. Only exposed efficiently seeded endogenous in cells. These findings suggest inhibiting enzymes mediating aggregates represents viable disease-modifying strategy slow progression other proteinopathies.

Language: Английский

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Development and validation of an expanded antibody toolset that captures alpha-synuclein pathological diversity in Lewy body diseases

npj Parkinson s Disease, Journal Year: 2023, Volume and Issue: 9(1)

Published: Dec. 7, 2023

The abnormal aggregation and accumulation of alpha-synuclein (aSyn) in the brain is a defining hallmark synucleinopathies. Various aSyn conformations post-translationally modified forms accumulate pathological inclusions vary abundance among these disorders. Relying on antibodies that have not been assessed for their ability to detect diverse may lead inaccurate estimations pathology human brains or disease models. To address this challenge, we developed characterized an expanded antibody panel targets different sequences post-translational modifications along length aSyn, recognizes all monomeric, oligomeric, fibrillar conformations. Next, profiled across sporadic familial Lewy body diseases (LBDs) reveal heterogeneous pathology, rich Serine 129 phosphorylation, Tyrosine 39 nitration N- C-terminal tyrosine phosphorylations, scattered both neurons glia. In addition, show can become hyperphosphorylated during processes inclusion maturation neuronal animal models seeding spreading. validation pipeline describe paves way systematic investigations into diversity brain, peripheral tissues, as well cellular

Language: Английский

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Brain-derived and in vitro-seeded alpha-synuclein fibrils exhibit distinct biophysical profiles

eLife, Journal Year: 2024, Volume and Issue: 13

Published: March 21, 2024

The alpha-synuclein (αSyn) seeding amplification assay (SAA) that allows the generation of disease-specific in vitro seeded fibrils (SAA fibrils) is used as a research tool to study connection between structure αSyn fibrils, cellular seeding/spreading, and clinicopathological manifestations different synucleinopathies. However, structural differences human brain-derived SAA have been recently highlighted. Here, we characterize biophysical properties from brains patients with Parkinson’s disease without dementia (PD, PDD), Lewy bodies (DLB), multiple system atrophy (MSA), compare them ‘model’ fibrils. We report show distinct biochemical profiles, which were not replicated corresponding Furthermore, all synucleinopathies displayed mixture ‘straight’ ‘twisted’ microscopic structures. PD, PDD, DLB had ’straight’ structure, whereas MSA showed structure. Finally, four phosphorylated (S129). Interestingly, carried over PDD fibrils. Our findings demonstrate limitation modeling pay attention necessity deepening understanding fibrillation methodology.

Language: Английский

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Brain-derived and in vitro-seeded alpha-synuclein fibrils exhibit distinct biophysical profiles

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Nov. 25, 2024

The alpha-synuclein (αSyn) seeding amplification assay (SAA) that allows the generation of disease-specific in vitro seeded fibrils (SAA fibrils) is used as a research tool to study connection between structure αSyn fibrils, cellular seeding/spreading, and clinicopathological manifestations different synucleinopathies. However, structural differences human brain-derived SAA have been recently highlighted. Here, we characterize biophysical properties from brains patients with Parkinson’s disease without dementia (PD, PDD), Lewy bodies (DLB), multiple system atrophy (MSA), compare them ‘model’ fibrils. We report show distinct biochemical profiles, which were not replicated corresponding Furthermore, all synucleinopathies displayed mixture ‘straight’ ‘twisted’ microscopic structures. PD, PDD, DLB had ’straight’ structure, whereas MSA showed structure. Finally, four phosphorylated (S129). Interestingly, carried over PDD fibrils. Our findings demonstrate limitation modeling pay attention necessity deepening understanding fibrillation methodology.

Language: Английский

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Fully co-factor-free ClearTau platform produces seeding-competent Tau fibrils for reconstructing pathological Tau aggregates

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: July 4, 2023

Tau protein fibrillization is implicated in the pathogenesis of several neurodegenerative diseases collectively known as Tauopathies. For decades, investigating vitro has required addition polyanions or other co-factors to induce its misfolding and aggregation, with heparin being most commonly used. However, heparin-induced fibrils exhibit high morphological heterogeneity a striking structural divergence from isolated Tauopathies patients' brains at ultra- macro-structural levels. To address these limitations, we developed quick, cheap, effective method for producing completely co-factor-free all full-length isoforms mixtures thereof. We show that generated using this ClearTau - amyloid-like features, possess seeding activity biosensor cells hiPSC-derived neurons, retain RNA-binding capacity, have properties structures more reminiscent brain-derived fibrils. present proof-of-concept implementation platform screening aggregation-modifying compounds. demonstrate advances open opportunities investigate pathophysiology disease-relevant aggregates will facilitate development pathology-targeting modifying therapies PET tracers can distinguish between different

Language: Английский

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Development and validation of an expanded antibody toolset that captures alpha-synuclein pathological diversity in Lewy body diseases

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: May 28, 2022

ABSTRACT The abnormal aggregation and accumulation of alpha-synuclein (aSyn) in the brain is a defining hallmark synucleinopathies. Different aSyn conformations post-translationally modified forms accumulate pathological inclusions vary abundance across Relying on antibodies that have not been assessed for their ability to capture diversity species may provide an accurate estimation pathology human brains or disease models. To address this challenge, we developed characterised expanded antibody panel targets different sequences post-translational modifications along length recognises all three (monomeric, oligomeric, fibrillar). Next, profiled sporadic familial Lewy body diseases (LBDs) reveal heterogeneously pathologies rich Serine 129 phosphorylation but also Tyrosine 39 nitration N- C-terminal tyrosine phosphorylations, scattered neurons glia. We show become hyperphosphorylated during inclusion maturation processes neuronal animal models spreading. validation pipeline describe here paves way more systematic investigations peripheral tissues, cellular

Language: Английский

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Brain-derived and in vitro-seeded alpha-synuclein fibrils exhibit distinct biophysical profiles

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 6, 2023

Abstract The alpha-synuclein ( α Syn) seeding amplification assay (SAA) that allows the generation of disease-specific in vitro seeded fibrils (SAA fibrils) is used as a research tool to study connection between structure Syn fibrils, cellular seeding/spreading, and clinico-pathological manifestations different synucleinopathies. However, structural differences human brain-derived SAA have been recently highlighted. Here, we characterize biophysical properties from brains patients with Parkinson’s disease without dementia (PD, PDD), Lewy bodies (DLB), multiple system atrophy (MSA) compare them ‘model’ fibrils. We report show distinct biochemical profiles, which were not replicated corresponding Furthermore, all synucleinopathies displayed mixture of- ‘straight’ ‘twisted’ microscopic structures. PD, PDD, DLB had structure, whereas MSA showed structure. Finally, four phosphorylated (S129). Interestingly, carried over PDD Our findings demonstrate limitation modelling pay attention necessity deepening understanding fibrillation methodology.

Language: Английский

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Combining non-canonical amino acid mutagenesis and native chemical ligation for multiply modifying proteins: A case study of α-synuclein post-translational modifications

Methods, Journal Year: 2023, Volume and Issue: 218, P. 101 - 109

Published: Aug. 6, 2023

Language: Английский

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