Author ORCID Identifier

https://orcid.org/0000-0002-0390-2689

Date of Graduation

5-2019

Document Type

Thesis (MS)

Program Affiliation

Biomedical Sciences

Degree Name

Masters of Science (MS)

Advisor/Committee Chair

Rodrigo Morales, Ph.D.

Committee Member

Darren Boehning, Ph.D.

Committee Member

Zheng Chen, Ph.D.

Committee Member

Kevin Morano, Ph.D.

Committee Member

Ines Moreno-Gonzalez, Ph.D.

Abstract

The misfolded prion protein causes and transmits disease in both humans and animals. As other infectious agents, prions display strain variation, which can generate different pathological outcomes in affected individuals. Unfortunately, there are no known therapies for these diseases, which at present are invariably fatal. In this work, the Protein Misfolding Cyclic Amplification technology (PMCA, an in vitro test that replicates minimum quantities of infectious prions) has been modified to screen for small molecules inhibiting prion protein misfolding in a strain-specific manner. In order to approach a high-throughput PMCA system, technical aspects in PMCA has been optimized for application of prions from laboratory rodents (i.e., mouse and Syrian hamsters) using a 96-well plate PMCA (96wp-PMCA) platform. Utilizing the 96wp-PMCA technique, a small number of anti-prion and anti-amyloid molecules has been tested against these prion strains using different solvents and at varying concentrations. My results show that regardless of sequence homology, prion strains are differentially responsive to known protein misfolding inhibitors. Continual optimization of PMCA towards a high-throughput system may be used not only for screening therapeutic agents but also for diagnosis.

Keywords

prions, protein misfolding cyclic amplification, small molecules, anti-prion, anti-amyloid

Available for download on Friday, May 08, 2020

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