Prions are infectious proteins that cause age-dependent neurodegeneration in humans and animals. The accumulation of an alternatively folded isoform of the prion protein (PrPSc) in the brains of humans and animals is the hallmark of the prion disorders that include Creutzfeldt-Jakob disease (CJD). The prion diseases are invariably fatal and no effective treatment exists. In this application, we propose to continue our studies directed toward devising effective medications for the treatment of CJD. Toward this goal, we want to identify several compounds that either alone or in combination substantially prolong the incubation times in transgenic mice. A model for our proposed research is reflected in our earlier work with bigenic mouse expressing an inducible PrP transgene. These studies showed a substantial prolongation of the incubation time by doxycycline-mediated inhibition of PrPc expression. Toward achieving this goal, we plan to adapt human (Hu) prion protein (PrP) based assays to the High Throughput Screening (HTS) of chemical libraries. Initially, we plan to continue using prion-infected mouse (Mo) neuroblastoma cells designated ScN2a for HTS studies. At the same time, we propose to use uninfected, human (Hu) glioblastoma cells in our HTS search for compounds that decrease HuPrPc levels. In addition, we propose to create cultured cells that propagate either wt Hu or chimeric Hu/Mo CJD prions. These novel cell lines will be used for HTS of chemical libraries. In the proposed studies, both protease-sensitive (s) and resistant (r) PrPSc will be measured in both dividing and non-dividing cells. Potent, non-toxic lead compounds with anti-prion activity will be chemically and computationally optimized by appropriate Structure-Activity-Relationship (SAR) studies. Some computational analyses will focus on the permeability of lead compounds across the bloodbrain- barrier (BBB) and other computational studies on the formation of PrPSc as well as structural models of this protein. Because of problems that plague prion-infected cultured cell assays, we propose to develop additional assays for HTS that include adaptation of an amyloid seeding assay (ASA) using HuPrPSc to seed the polymerization of recombinant (rec) HuPrP as well as a humanized version of a fluorescence polarization (FP) assay that measures the binding of PrP peptides to HuPrPSc. We also propose to use transgenic (Tg) mice expressing either a chimeric Hu/Mo PrP or wt HuPrP that are susceptible to Hu CJD prions. One line of Tg mice exhibits incubation times of ~80 days and these animals should greatly facilitate our studies.

Public Health Relevance

The ultimate goal of these studies is to develop drugs that cure human prion disorders including CJD. The successful development of a therapeutic for CJD would have important implications for creating meaningful treatments for other age-dependent neurodegenerative diseases that include Alzheimer's disease.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Program Projects (P01)
Project #
5P01AG021601-09
Application #
8231346
Study Section
Special Emphasis Panel (ZAG1-ZIJ-7 (J1))
Program Officer
Mackiewicz, Miroslaw
Project Start
2003-06-01
Project End
2014-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
9
Fiscal Year
2012
Total Cost
$1,893,873
Indirect Cost
$668,065
Name
University of California San Francisco
Department
Neurology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Watts, Joel C; Giles, Kurt; Saltzberg, Daniel J et al. (2016) Guinea Pig Prion Protein Supports Rapid Propagation of Bovine Spongiform Encephalopathy and Variant Creutzfeldt-Jakob Disease Prions. J Virol 90:9558-9569
Watts, Joel C; Giles, Kurt; Bourkas, Matthew E C et al. (2016) Towards authentic transgenic mouse models of heritable PrP prion diseases. Acta Neuropathol 132:593-610
Ahlenius, Henrik; Chanda, Soham; Webb, Ashley E et al. (2016) FoxO3 regulates neuronal reprogramming of cells from postnatal and aging mice. Proc Natl Acad Sci U S A 113:8514-9
Geschwind, Michael D (2016) Rapidly Progressive Dementia. Continuum (Minneap Minn) 22:510-37
Giles, Kurt; Berry, David B; Condello, Carlo et al. (2016) Optimization of Aryl Amides that Extend Survival in Prion-Infected Mice. J Pharmacol Exp Ther 358:537-47
Elkins, Matthew R; Wang, Tuo; Nick, Mimi et al. (2016) Structural Polymorphism of Alzheimer's β-Amyloid Fibrils as Controlled by an E22 Switch: A Solid-State NMR Study. J Am Chem Soc 138:9840-52
Giles, Kurt; Berry, David B; Condello, Carlo et al. (2015) Different 2-Aminothiazole Therapeutics Produce Distinct Patterns of Scrapie Prion Neuropathology in Mouse Brains. J Pharmacol Exp Ther 355:2-12
Kim, Mee-Ohk; Geschwind, Michael D (2015) Clinical update of Jakob-Creutzfeldt disease. Curr Opin Neurol 28:302-10
Ng, Adeline S L; Kramer, Joel; Centurion, Alejandro et al. (2015) Clinico-pathological correlation in adenylate kinase 5 autoimmune limbic encephalitis. J Neuroimmunol 287:31-5
Lu, Jonathan; Trnka, Michael J; Roh, Soung-Hun et al. (2015) Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes. J Am Soc Mass Spectrom 26:2141-51

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