PrioNet Co-Funded Projects

Dr. Carmen Fuentealba, University of Calgary &

Dr. Stéphane McLachlan, University of Manitoba

“In Land and Life: Multi-Scale Implications of Chronic Wasting Disease for Aboriginal Communities  and Other Stakeholders”

Chronic wasting disease (CWD) is found in deer, elk, and moose across western US and Canada. It may have severe socioeconomic and health implications for hunters, the nature industry, and Aboriginal communities. Although widely recognized as a research and management priority, no published study has characterized the vulnerability of these communities to CWD.  Our multidisciplinary study will cross biological, socioeconomic and cultural dimensions of CWD and will be conducted at local, regional, national, and international scales of organization in collaboration with Aboriginal communities in Alberta. It will address severe gaps in communication regarding CWD and other wildlife disease that have implications for human and environmental health.

Our specific objectives are to: 1) Characterize the hunting and consumption of cervids in Aboriginal communities within central Alberta; 2) Assess concerns of Aboriginal communities and organizations relative to other stakeholders (i.e. hunters, tourism, ranchers, scientists, agencies etc.) regarding CWD and other wildlife disease; 3) Better understand the mechanisms underlying any impacts; and 4) Develop diverse forms of communication in collaboration with these communities and stakeholders that better inform risks associated with CWD for Aboriginal peoples, decision-makers, and the public as a whole.

“Characterizing Patterns of Contagion and Transmission of Chronic Wasting Disease in Cervids”

Critical knowledge gaps exist in our understandings about the transmission of CWD, and include questions such as: i) when does an infected animal begin shedding prions and become contagious to other animals; ii) which tissues or body fluids are responsible for contagion, iii) what is the burden of prion contamination in these fluids and tissues; iv) do animals become infected through direct animal to animal contact (e.g., grooming or other social interactions) and/or indirectly through the environments where prions can accumulate to the doses necessary establish infections (i.e., over wintering grounds, high density food sources). We propose to use protein misfolding cyclic amplification (PMCA) assays to characterize patterns of transmission and contagion in infected animals (white-tailed deer) and apply these tools to surveillance of captive and free-ranging animals. This basic knowledge will be critical in developing management strategies aimed at controlling the spread of the disease, and minimizing potential impacts to both human and animal health. 

“Oligomeric PrPres and amyloidogenic PrP fragments: their molecular structure, toxicity, and role in prion disease ”

Protease-resistant prion protein (PrPres) is the major protein component of the infectious agent in prion diseases. How pathogenesis develops is unclear, but it is strongly influenced by the conformation of PrPres, the size of PrPres oligomer1, and its interaction with membranes2. This project will examine these three aspects of pathogenesis using a multi-disciplinary approach combining organotypic culture assays, microarray analyses, solid state NMR, and computer simulations.   Dr. Sim's research team will study the structure of small portions of the PrP protein.  The toxicity of different sized clusters of PrP protein will be measured.  This will help us better understand how the disease progresses in the brain.

 Protein kinases are enzymes that play a role in the development of most infectious and neurodegenerative diseases, they are important targets for drugs used to treat these diseases. However, little is known about their role in prion diseases.  Dr. Schang plans to use targeted proteomics (molecular studies of large groups of proteins in cells) to identify the specific protein kinases involved in the development of prion diseases and the role these enzymes play in the spread of prion infection. Understanding the molecular basis of prion diseases may help scientists develop new drugs that can slow down or prevent these diseases.

“Protein folding diseases in zebrafish”

The aim of this proposal is the creation of genetically altered,   zebrafish that will represent a potent animal model of Prion Diseases and other abnormal protein development diseases. The emerging usefulness of zebrafish is rooted in the ability to efficiently alter gene function. Rapid external development combined with the ease of inexpensively producing many embryos; make this a quick and affordable system for examining various prion experiments. .The zebrafish can live for 3-5 years, allowing time for the slow development of neuropathologies. The zebrafish is also amenable to behavioural and electrophysiological experimentation. Overall the system combines genetic and developmental tractability of genes. 

The project will analyze the role of PrP and amyloids in neuroprotection and adult neurogenesis. The mutant and transgenic zebrafish created in this proposal, combined with the large community of zebrafish reagents and resources, will be a powerful platform to quickly analyze the role of proteins that interact with PrP and for primary screens of small molecule inhibitors. 

The central objective for this project is to develop inexpensive, non-invasive test for diagnosing and monitoring prion diseases (CJD, vCJD, BSE, CWD and other TSE's) using non-prion biomarkers. To this end we intend to apply novel, quantitative metabolomic and proteomic techniques that have recently been developed in our laboratories towards the identification and validation of preclinical biomarkers for prion diseases. In particular we will focus on finding novel and confirming supposed prion disease biomarkers in blood, urine, feces and Cerebrospinal fluid (CSF) using animal models. The team will use high resolution mass spectroscopy methods to identify and quantify large numbers of metabolites and protein biomarkers. These novel methods of detecting biomarkers have a higher sensitivity than have been used in previous prion studies. The long-term goal for this project is to develop an inexpensive and non-invasive test that can be quickly done on live animals or live individuals prior to the onset of disease symptoms or transmitability. Such a test would allow inexpensive monitoring of BSE in cattle herds, simplify the tracking and mitigation of CWD in wild cervid populations and improve treatment or intervention options for CJD or vCJD affected humans.

Chronic Wasting Disease (CWD) and Scrapie are endemic TSE diseases in North America. CWD and scrapie share some key characteristics, the most important being that the infectious agent in both of these diseases can be horizontally transmitted and can lead to environmental contamination. Understanding the process of infection, disease progression, and detecting transmission at an early stage are vital to controlling these diseases.

Approved diagnostic methods for CWD and scrapie are based on post mortem identification of disease-associated prion proteins. The limitation to these methods means that detection of a single positive animal forces control programs to depopulate animals within a certain radius of the index case, despite the fact they may be perfectly healthy. In the project, they will approach the problems of late diagnosis and lack of disease understanding using gene expression analysis. The project will identify and adopt technologies that will serve to improve our understanding of TSE diseases and their interactions with their hosts. This improved understanding could lead to more efficient and effective diagnostic, control, and eradication programs and reduce the impact of disease outbreaks.

Prion proteins are natural and relatively harmless proteins found in the bodies of humans and almost all animals; however, prion proteins are quite unique in that they can change shape and thereby "morph" from a harmless protein to a harmful protein complex. A key question that is still unanswered is: what is the shape or conformation of this harmful form of the prion protein? This project uses a technique called mass spectrometry in combination with specialized chemical modifications to help figure out what this harmful version of the prion protein actually looks like.

Determining the structure of this prion protein complex could lead to new ways to distinguish between the "healthy" and the "harmful" version of the prion protein. This can possibly lead to new ways that prevent the harmful version forming in the first place.