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Prolyl oligopeptidase inhibition by KYP-2407 increases alpha-synuclein fibril degradation in neuron-like cells

Rostami, Jinar ; Jäntti, Maria ; Cui, Hengjing ; Rinne, Maiju K ; Kukkonen, Jyrki P ; Falk, Anna LU ; Erlandsson, Anna and Myöhänen, Timo (2020) In Biomedicine and Pharmacotherapy 131.
Abstract

Growing evidence emphasizes insufficient clearance of pathological alpha-synuclein (αSYN) aggregates in the progression of Parkinson's disease (PD). Consequently, cellular degradation pathways represent a potential therapeutic target. Prolyl oligopeptidase (PREP) is highly expressed in the brain and has been suggested to increase αSYN aggregation and negatively regulate the autophagy pathway. Inhibition of PREP with a small molecule inhibitor, KYP-2407, stimulates autophagy and reduces the oligomeric species of αSYN aggregates in PD mouse models. However, whether PREP inhibition has any effects on intracellular αSYN fibrils has not been studied before. In this study, the effect of KYP2407 on αSYN preformed fibrils (PFFs) was tested in... (More)

Growing evidence emphasizes insufficient clearance of pathological alpha-synuclein (αSYN) aggregates in the progression of Parkinson's disease (PD). Consequently, cellular degradation pathways represent a potential therapeutic target. Prolyl oligopeptidase (PREP) is highly expressed in the brain and has been suggested to increase αSYN aggregation and negatively regulate the autophagy pathway. Inhibition of PREP with a small molecule inhibitor, KYP-2407, stimulates autophagy and reduces the oligomeric species of αSYN aggregates in PD mouse models. However, whether PREP inhibition has any effects on intracellular αSYN fibrils has not been studied before. In this study, the effect of KYP2407 on αSYN preformed fibrils (PFFs) was tested in SH-SY5Y cells and human astrocytes. Immunostaining analysis revealed that both cell types accumulated αSYN PFFs intracellularly but KYP-2047 decreased intracellular αSYN deposits only in SH-SY5Y cells, as astrocytes did not show any PREP activity. Western blot analysis confirmed the reduction of high molecular weight αSYN species in SH-SY5Y cell lysates, and secretion of αSYN from SH-SY5Y cells also decreased in the presence of KYP-2407. Accumulation of αSYN inside the SH-SY5Y cells resulted in an increase of the auto-lysosomal proteins p62 and LC3BII, as well as calpain 1 and 2, which have been shown to be associated with PD pathology. Notably, treatment with KYP-2407 significantly reduced p62 and LC3BII levels, indicating an increased autophagic flux, and calpain 1 and 2 levels returned to normal in the presence of KYP-2407. Our findings indicate that PREP inhibition can potentially be used as therapy to reduce the insoluble intracellular αSYN aggregates.

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Astrocytes/drug effects, Autophagy/drug effects, Cell Line, Tumor, Cells, Cultured, Disease Progression, Humans, Neurons/drug effects, Parkinson Disease/physiopathology, Proline/analogs & derivatives, Prolyl Oligopeptidases/antagonists & inhibitors, Serine Proteinase Inhibitors/pharmacology, alpha-Synuclein/metabolism
in
Biomedicine and Pharmacotherapy
volume
131
article number
110788
pages
10 pages
publisher
Elsevier
external identifiers
  • scopus:85091580966
  • pmid:33152946
ISSN
1950-6007
DOI
10.1016/j.biopha.2020.110788
language
English
LU publication?
no
id
9c2c92c4-e133-48df-96aa-4a902eb9deab
date added to LUP
2021-08-09 13:43:40
date last changed
2024-06-15 13:54:25
@article{9c2c92c4-e133-48df-96aa-4a902eb9deab,
  abstract     = {{<p>Growing evidence emphasizes insufficient clearance of pathological alpha-synuclein (αSYN) aggregates in the progression of Parkinson's disease (PD). Consequently, cellular degradation pathways represent a potential therapeutic target. Prolyl oligopeptidase (PREP) is highly expressed in the brain and has been suggested to increase αSYN aggregation and negatively regulate the autophagy pathway. Inhibition of PREP with a small molecule inhibitor, KYP-2407, stimulates autophagy and reduces the oligomeric species of αSYN aggregates in PD mouse models. However, whether PREP inhibition has any effects on intracellular αSYN fibrils has not been studied before. In this study, the effect of KYP2407 on αSYN preformed fibrils (PFFs) was tested in SH-SY5Y cells and human astrocytes. Immunostaining analysis revealed that both cell types accumulated αSYN PFFs intracellularly but KYP-2047 decreased intracellular αSYN deposits only in SH-SY5Y cells, as astrocytes did not show any PREP activity. Western blot analysis confirmed the reduction of high molecular weight αSYN species in SH-SY5Y cell lysates, and secretion of αSYN from SH-SY5Y cells also decreased in the presence of KYP-2407. Accumulation of αSYN inside the SH-SY5Y cells resulted in an increase of the auto-lysosomal proteins p62 and LC3BII, as well as calpain 1 and 2, which have been shown to be associated with PD pathology. Notably, treatment with KYP-2407 significantly reduced p62 and LC3BII levels, indicating an increased autophagic flux, and calpain 1 and 2 levels returned to normal in the presence of KYP-2407. Our findings indicate that PREP inhibition can potentially be used as therapy to reduce the insoluble intracellular αSYN aggregates.</p>}},
  author       = {{Rostami, Jinar and Jäntti, Maria and Cui, Hengjing and Rinne, Maiju K and Kukkonen, Jyrki P and Falk, Anna and Erlandsson, Anna and Myöhänen, Timo}},
  issn         = {{1950-6007}},
  keywords     = {{Astrocytes/drug effects; Autophagy/drug effects; Cell Line, Tumor; Cells, Cultured; Disease Progression; Humans; Neurons/drug effects; Parkinson Disease/physiopathology; Proline/analogs & derivatives; Prolyl Oligopeptidases/antagonists & inhibitors; Serine Proteinase Inhibitors/pharmacology; alpha-Synuclein/metabolism}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{Biomedicine and Pharmacotherapy}},
  title        = {{Prolyl oligopeptidase inhibition by KYP-2407 increases alpha-synuclein fibril degradation in neuron-like cells}},
  url          = {{https://lup.lub.lu.se/search/files/101030811/Prolyl_oligopeptidase_inhibition.pdf}},
  doi          = {{10.1016/j.biopha.2020.110788}},
  volume       = {{131}},
  year         = {{2020}},
}