Screening of Anti-Prion Compounds Using the Protein Misfolding Cyclic Amplification Technology

Authors

Sandra Pritzkow
Isaac Schauer
Ananya Tupaki-Sreepurna
Rodrigo Morales
Claudio Soto

Publication Date

9-4-2024

Journal

Biomolecules

Abstract

Prion diseases are 100% fatal infectious neurodegenerative diseases affecting the brains of humans and other mammals. The disease is caused by the formation and replication of prions, composed exclusively of the misfolded prion protein (PrPSc). We invented and developed the protein misfolding cyclic amplification (PMCA) technology for in vitro prion replication, which allow us to replicate the infectious agent and it is commonly used for ultra-sensitive prion detection in biological fluids, tissues and environmental samples. In this article, we studied whether PMCA can be used to screen for chemical compounds that block prion replication. A small set of compounds previously shown to have anti-prion activity in various systems, mostly using cells infected with murine prions, was evaluated for their ability to prevent the replication of prions. Studies were conducted simultaneously with prions derived from 4 species, including human, cattle, cervid and mouse. Our results show that only one of these compounds (methylene blue) was able to completely inhibit prion replication in all species. Estimation of the IC50 for methylene blue inhibition of human prions causing variant Creutzfeldt-Jakob disease (vCJD) was 7.7 μM. Finally, we showed that PMCA can be used for structure-activity relationship studies of anti-prion compounds. Interestingly, some of the less efficient prion inhibitors altered the replication of prions in some species and not others, suggesting that PMCA is useful for studying the differential selectivity of potential drugs.

Keywords

Animals, Humans, Mice, Protein Folding, Cattle, Methylene Blue, Prion Diseases, Prions, PrPSc Proteins, Drug Evaluation, Preclinical

Comments

Supplementary Materials

Data Availability Statement

PMID: 39334879