Prion diseases are infectious and fatal neurodegenerative disorders with no available treatment. Zoonotic prion transmission is of concern, as the bovine spongiform encephalopathy (BSE) epidemic has led to nearly 200 cases of prion infection in humans. New prion diseases have recently emerged in food animals and wild deer with unknown potential for spread to humans. The molecular mechanisms that underlie prion aggregation, transmission between species, and the prion strains are still poorly understood. Our long term goal is to identify the key residues of the prion protein that govern species barriers and strain conformation. The ?2-a2 loop of the prion protein (amino acids 165-175) is a site of exceptionally high sequence variability. Microcrystallography has shown that adjacent ?2-a2 loops can align as ?-sheets with side chains that intermesh in a dry interface as part of the amyloid core. We have shown that two amino acid substitutions in the ?2-a2 loop of the mouse prion protein lead to de novo prion disease in transgenic mice. In preliminary studies, these substitutions markedly change the interspecies transmission barriers in mice. Here we propose that the ?2-a2 loop also plays an important role for prion transmission to humans and cattle. In this proposal, we will evaluate the impact of the ?2-a2 loop residues on prion aggregation, species barriers, and strains.
In Aim 1, we will determine the role of critical residues for prion aggregation and conformational conversion in vitro and in vivo.
In Aim 2, we will assess the importance of the ?2-a2 loop region for prion infection of humans and cattle using mouse models that differ only at the loop. Results from these studies will contribute to our understanding of the fundamental mechanisms of prion protein aggregation, aid in our risk assessment of prion transmission to humans, as well as advance the rational design of therapeutics to block prion aggregation.

Public Health Relevance

Prion infections have the potential to spread from animals to humans and have caused deaths as well as enormous economic losses in the UK. Chronic wasting disease (CWD) is an emerging prion disease in deer and elk, transmitted at a rate unparalleled by any other prion disease, and thus may present a risk of infecting humans, our food animals, and other wildlife. We propose to investigate the underlying mechanisms for when and how prions can infect a new host.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS069566-02
Application #
8240997
Study Section
Clinical Neuroimmunology and Brain Tumors Study Section (CNBT)
Program Officer
Wong, May
Project Start
2011-03-15
Project End
2016-02-29
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
2
Fiscal Year
2012
Total Cost
$304,887
Indirect Cost
$91,696
Name
University of California San Diego
Department
Pathology
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Kurt, Timothy D; Sigurdson, Christina J (2016) Cross-species transmission of CWD prions. Prion 10:83-91
Kurt, Timothy D; Jiang, Lin; Fernández-Borges, Natalia et al. (2015) Human prion protein sequence elements impede cross-species chronic wasting disease transmission. J Clin Invest 125:1485-96
Hiramatsu, Nobuhiko; Chiang, Wei-Chieh; Kurt, Timothy D et al. (2015) Multiple Mechanisms of Unfolded Protein Response-Induced Cell Death. Am J Pathol 185:1800-8
Kurt, Timothy D; Jiang, Lin; Fernández-Borges, Natalia et al. (2015) Human prion protein sequence elements impede cross-species chronic wasting disease transmission. J Clin Invest 125:2548
Joshi-Barr, Shivanjali; Bett, Cyrus; Chiang, Wei-Chieh et al. (2014) De novo prion aggregates trigger autophagy in skeletal muscle. J Virol 88:2071-82
Kurt, Timothy D; Jiang, Lin; Bett, Cyrus et al. (2014) A proposed mechanism for the promotion of prion conversion involving a strictly conserved tyrosine residue in the β2-α2 loop of PrPC. J Biol Chem 289:10660-7
Magnusson, Karin; Simon, Rozalyn; Sjölander, Daniel et al. (2014) Multimodal fluorescence microscopy of prion strain specific PrP deposits stained by thiophene-based amyloid ligands. Prion 8:319-29
Kurt, Timothy D; Bett, Cyrus; Fernández-Borges, Natalia et al. (2014) Prion transmission prevented by modifying the β2-α2 loop structure of host PrPC. J Neurosci 34:1022-7
Klingstedt, Therése; Shirani, Hamid; Åslund, K O Andreas et al. (2013) The structural basis for optimal performance of oligothiophene-based fluorescent amyloid ligands: conformational flexibility is essential for spectral assignment of a diversity of protein aggregates. Chemistry 19:10179-92
Bett, Cyrus; Kurt, Tim D; Lucero, Melanie et al. (2013) Defining the conformational features of anchorless, poorly neuroinvasive prions. PLoS Pathog 9:e1003280

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