Use of the Protein Misfolding Cyclic Amplification for food safety and drug discovery

Author

Maria R. Benavente Garcia-Huidobro, The University of Texas MD Anderson Cancer CenterFollow

Author ORCID Identifier

0009-0000-0271-9674

Date of Graduation

8-2025

Document Type

Thesis (MS)

Program Affiliation

Neuroscience

Degree Name

Masters of Science (MS)

Advisor/Committee Chair

Rodrigo Morales

Committee Member

Seung-Hee Yoo

Committee Member

Kartik Venkatachalam

Committee Member

Sheng Zhang

Committee Member

Tatiana Barichello

Abstract

Prion diseases are fatal neurodegenerative disorders caused by the misfolding of the normal prion protein (PrP^C) into its infectious form (PrP^Sc). While the zoonotic potential of chronic wasting disease (CWD) remains uncertain, the presence of prions in food products raises public health concerns. Additionally, therapeutic strategies for prion diseases remain limited. This thesis presents a methodological exploration of the Protein Misfolding Cyclic Amplification (PMCA) technique to address these challenges in two key areas: (1) detecting CWD prions in processed meats subjected to common cooking procedures, and (2) identifying potential anti-prion compounds through a high-throughput PMCA adaptation.

Our results demonstrate that PMCA effectively detected CWD prions in a range of processed meat products, with an unexpected increase in prion detectability following grilling and boiling, suggesting enhanced prion accessibility post-cooking. Despite this, these prions failed to convert the human prion protein in PMCA reactions, indicating a limited zoonotic risk under the conditions/materials tested.

In parallel, a modified 96-well plate PMCA screening strategy identified eight promising compounds with inhibitory effects on PrP^Sc formation. Notably, these candidates exhibited diverse properties, including both hydrophilic and hydrophobic profiles, with some compounds previously linked to amyloidogenic pathway inhibition.

This study highlights the versatility of the PMCA technique for both, food safety assessments and screening anti-prion compounds. The results presented in this thesis work stress the importance of exploring prion strain diversity, refining PMCA protocols, and validating potential inhibitors in in vivo models for future therapeutic development.

Keywords

Prion, drug discovery, food safety, Protein misfolding cyclic amplification, Chronic wasting disease, zoonosis