Prion diseases, such as Creutzfeldt-Jakob Disease (CJD), are degenerative disorders affecting the central nervous system. They are uniformly fatal and illicit death following a long period of steady neurodegeneration, dementia and motor disfunction. In prion diseases, an aberrantly folded conformer ofthe prion protein (PrPSc) propagates by catalyzing the post-translational misfolding of cellular PrP (PrPC). We have recently succeeded in recapitulating this phenomenon in vitro by converting purified, bacterially-expressed, recombinant PrP into infectious prions. The ability to create synthetic prions in cell-free systems has provided definitive proof of the protein-only hypothesis of prion propagation and has enabled us to begin deciphering the structural basis of protein-based infectivity.
The specific aims of this project seek to utilize synthetic prions to gain insights into the mechanism of prion-induced neurodegeneration. We propose to create homogeneous and highly infectious preparations of synthetic prions. The synthetic prion preparations will be propagated in mouse bioassays and used for structural analyses. Disease phenotypes induced by synthetic prions will be assessed by two novel approaches for global analysis of proteome homeostasis. Effects on protein translation will be studied by ribosome profiling and changes in proteome turnover will be measured by comprehensive in vivo isotopic replacement. We will also attempt to infect induced neuronal (iN] cells with synthetic prions. These strategies will uncover relationships between the physical structure of prion strains and their biological properties and may provide valuable insights into the mechanism of neurodegeneration in prion diseases such as CJD.

Public Health Relevance

The continuing aging ofthe American population has made the discovery of therapies against neurodegenerative disorders an important public health goal. This study will provide insight into the etiology of prion diseases such as Creutzfeldt-Jakob disease (CJD), and may illuminate the mechanism of neurodegeneration in more prevalent proteinopathies such as Alzheimer's disease and Parkinson's disease.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Program Projects (P01)
Project #
5P01AG010770-20
Application #
8448174
Study Section
Special Emphasis Panel (ZAG1-ZIJ-6)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
20
Fiscal Year
2013
Total Cost
$357,131
Indirect Cost
$127,045
Name
University of California San Francisco
Department
Type
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Watts, Joel C; Giles, Kurt; Patel, Smita et al. (2014) Evidence that bank vole PrP is a universal acceptor for prions. PLoS Pathog 10:e1003990
Watts, Joel C; Prusiner, Stanley B (2014) Mouse models for studying the formation and propagation of prions. J Biol Chem 289:19841-9
Watts, Joel C; Condello, Carlo; Stöhr, Jan et al. (2014) Serial propagation of distinct strains of A? prions from Alzheimer's disease patients. Proc Natl Acad Sci U S A 111:10323-8
Burré, Jacqueline; Sharma, Manu; Südhof, Thomas C (2014) ?-Synuclein assembles into higher-order multimers upon membrane binding to promote SNARE complex formation. Proc Natl Acad Sci U S A 111:E4274-83
Chanda, Soham; Ang, Cheen Euong; Davila, Jonathan et al. (2014) Generation of induced neuronal cells by the single reprogramming factor ASCL1. Stem Cell Reports 3:282-96
Ang, Cheen Euong; Wernig, Marius (2014) Induced neuronal reprogramming. J Comp Neurol 522:2877-86
Silber, B Michael; Gever, Joel R; Rao, Satish et al. (2014) Novel compounds lowering the cellular isoform of the human prion protein in cultured human cells. Bioorg Med Chem 22:1960-72
Stöhr, Jan; Condello, Carlo; Watts, Joel C et al. (2014) Distinct synthetic A? prion strains producing different amyloid deposits in bigenic mice. Proc Natl Acad Sci U S A 111:10329-34
Li, Zhe; Gever, Joel; Rao, Satish et al. (2013) Discovery and Preliminary SAR of Arylpiperazines as Novel, Brainpenetrant Antiprion Compounds. ACS Med Chem Lett 4:397-401
Berry, David B; Lu, Duo; Geva, Michal et al. (2013) Drug resistance confounding prion therapeutics. Proc Natl Acad Sci U S A 110:E4160-9

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