Identification of compounds inhibiting prion replication and toxicity by removing PrPC from the cell surface

Silvia Biggi, Michael Pancher, Claudia Stincardini, Silvia Luotti, Tania Massignan, Andrea Dalle Vedove, Andrea Astolfi, Pamela Gatto, Graziano Lolli, Maria Letizia Barreca, Valentina Bonetto, Valentina Adami, Emiliano Biasini

Research output: Contribution to journalArticlepeer-review

Abstract

The vast majority of therapeutic approaches tested so far for prion diseases, transmissible neurodegenerative disorders of human and animals, tackled PrPSc , the aggregated and infectious isoform of the cellular prion protein (PrPC ), with largely unsuccessful results. Conversely, targeting PrPC expression, stability or cell surface localization are poorly explored strategies. We recently characterized the mode of action of chlorpromazine, an anti-psychotic drug known to inhibit prion replication and toxicity by inducing the re-localization of PrPC from the plasma membrane. Unfortunately, chlorpromazine possesses pharmacokinetic properties unsuitable for chronic use in vivo, namely low specificity and high toxicity. Here, we employed HEK293 cells stably expressing EGFP-PrP to carry out a semi-automated high content screening (HCS) of a chemical library directed at identifying non-cytotoxic molecules capable of specifically relocalizing PrPC from the plasma membrane as well as inhibiting prion replication in N2a cell cultures. We identified four candidate hits inducing a significant reduction in cell surface PrPC , one of which also inhibited prion propagation and toxicity in cell cultures in a strain-independent fashion. This study defines a new screening method and novel anti-prion compounds supporting the notion that removing PrPC from the cell surface could represent a viable therapeutic strategy for prion diseases.

Original languageEnglish
Pages (from-to)136-150
Number of pages15
JournalJournal of Neurochemistry
Volume152
Issue number1
DOIs
Publication statusPublished - Jan 2020

Keywords

  • Animals
  • Casein Kinase II/antagonists & inhibitors
  • Cell Line, Tumor
  • Cell Membrane/chemistry
  • Cell Survival/drug effects
  • Drug Evaluation, Preclinical/methods
  • Fluorescent Dyes
  • Gene Expression
  • Green Fluorescent Proteins/analysis
  • HEK293 Cells
  • Harmaline/analogs & derivatives
  • Hematoxylin/analogs & derivatives
  • Humans
  • Mice
  • Neuroblastoma
  • PrPC Proteins/analysis
  • Prions/antagonists & inhibitors
  • Quinacrine/pharmacology
  • Tacrolimus/pharmacology

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