FRET-Based Screening Identifies p38 MAPK and PKC Inhibition as Targets for Prevention of Seeded α-Synuclein Aggregation. Academic Article uri icon

Overview

abstract

  • 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.

publication date

  • July 13, 2021

Research

keywords

  • Enzyme Inhibitors
  • Fluorescence Resonance Energy Transfer
  • Protein Aggregation, Pathological
  • Protein Kinase C
  • alpha-Synuclein
  • p38 Mitogen-Activated Protein Kinases

Identity

PubMed Central ID

  • PMC8609038

Scopus Document Identifier

  • 85110712889

Digital Object Identifier (DOI)

  • 10.1007/s13311-021-01070-1

PubMed ID

  • 34258749

Additional Document Info

volume

  • 18

issue

  • 3