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FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation
Aggregation of α-synuclein is associated with neurodegeneration and a hallmark pathology in synucleinopathies. These aggregates are thought to function as prion-like particles where the conformation of misfolded α-synuclein determines the traits of the induced pathology, similar to prion diseases. S...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8609038/ https://www.ncbi.nlm.nih.gov/pubmed/34258749 http://dx.doi.org/10.1007/s13311-021-01070-1 |
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author | Svanbergsson, Alexander Ek, Fredrik Martinsson, Isak Rodo, Jordi Liu, Di Brandi, Edoardo Haikal, Caroline Torres-Garcia, Laura Li, Wen Gouras, Gunnar Olsson, Roger Björklund, Tomas Li, Jia-Yi |
author_facet | Svanbergsson, Alexander Ek, Fredrik Martinsson, Isak Rodo, Jordi Liu, Di Brandi, Edoardo Haikal, Caroline Torres-Garcia, Laura Li, Wen Gouras, Gunnar Olsson, Roger Björklund, Tomas Li, Jia-Yi |
author_sort | Svanbergsson, Alexander |
collection | PubMed |
description | Aggregation of α-synuclein is associated with neurodegeneration and a hallmark pathology in synucleinopathies. These aggregates are thought to function as prion-like particles where the conformation of misfolded α-synuclein determines the traits of the induced pathology, similar to prion diseases. Still, little is known about the molecular targets facilitating the conformation-specific biological effects, but their identification could form the basis for new therapeutic interventions. High-throughput screening of annotated compound libraries could facilitate mechanistic investigation by identifying targets with impact on α-synuclein aggregation. To this end, we developed a FRET-based cellular reporter in HEK293T cells, with sensitivity down to 6.5 nM α-synuclein seeds. Using this model system, we identified GF109203X, SB202190, and SB203580 as inhibitors capable of preventing induction of α-synuclein aggregation via inhibition of p38 MAPK and PKC, respectively. We further investigated the mechanisms underlying the protective effects and found alterations in the endo-lysosomal system to be likely candidates of the protection. We found the changes did not stem from a reduction in uptake but rather alteration of lysosomal abundance and degradative capacity. Our findings highlight the value high-throughput screening brings to the mechanistic investigation of α-synuclein aggregation while simultaneously identifying novel therapeutic compounds. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13311-021-01070-1. |
format | Online Article Text |
id | pubmed-8609038 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-86090382021-12-03 FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation Svanbergsson, Alexander Ek, Fredrik Martinsson, Isak Rodo, Jordi Liu, Di Brandi, Edoardo Haikal, Caroline Torres-Garcia, Laura Li, Wen Gouras, Gunnar Olsson, Roger Björklund, Tomas Li, Jia-Yi Neurotherapeutics Original Article Aggregation of α-synuclein is associated with neurodegeneration and a hallmark pathology in synucleinopathies. These aggregates are thought to function as prion-like particles where the conformation of misfolded α-synuclein determines the traits of the induced pathology, similar to prion diseases. Still, little is known about the molecular targets facilitating the conformation-specific biological effects, but their identification could form the basis for new therapeutic interventions. High-throughput screening of annotated compound libraries could facilitate mechanistic investigation by identifying targets with impact on α-synuclein aggregation. To this end, we developed a FRET-based cellular reporter in HEK293T cells, with sensitivity down to 6.5 nM α-synuclein seeds. Using this model system, we identified GF109203X, SB202190, and SB203580 as inhibitors capable of preventing induction of α-synuclein aggregation via inhibition of p38 MAPK and PKC, respectively. We further investigated the mechanisms underlying the protective effects and found alterations in the endo-lysosomal system to be likely candidates of the protection. We found the changes did not stem from a reduction in uptake but rather alteration of lysosomal abundance and degradative capacity. Our findings highlight the value high-throughput screening brings to the mechanistic investigation of α-synuclein aggregation while simultaneously identifying novel therapeutic compounds. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13311-021-01070-1. Springer International Publishing 2021-07-13 2021-07 /pmc/articles/PMC8609038/ /pubmed/34258749 http://dx.doi.org/10.1007/s13311-021-01070-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Original Article Svanbergsson, Alexander Ek, Fredrik Martinsson, Isak Rodo, Jordi Liu, Di Brandi, Edoardo Haikal, Caroline Torres-Garcia, Laura Li, Wen Gouras, Gunnar Olsson, Roger Björklund, Tomas Li, Jia-Yi FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title | FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title_full | FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title_fullStr | FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title_full_unstemmed | FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title_short | FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation |
title_sort | fret-based screening identifies p38 mapk and pkc inhibition as targets for prevention of seeded α-synuclein aggregation |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8609038/ https://www.ncbi.nlm.nih.gov/pubmed/34258749 http://dx.doi.org/10.1007/s13311-021-01070-1 |
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