• Where is PRP(c) expressed?  Which tissues would antisense need to target?  Which symptoms are mediated by neurological damage v. caused by PrP(Sc) in other tissues?  Can we learn more about this from raw data produced by microarray studies?
  • What, if anything, can we learn about protein structure and function and possible therapeutic targets from multiple examples of dominant negative inhibition on the PRNP gene?  129 M/V; G127V (Kuru); E219K (Japan)
  • Why do multiple strains of prion disease target different areas of the brain – even when transferred into mice with normal genotypes?  Are particular types of aggregates more toxic to particular parts of the brain?  Are slightly different oligomers more prone to attack/ interact with different tissues?
  • Why does FFI target the thalamus?  What else do we know about the thalamus?
  • Which proteins interact with PrP(c)? (“interactome”)
  • Splicing of PRNP – In addition to PrP(c), does this gene produce other, minor forms of the protein if spliced differently?  If so, what role might these play in disease course?  (Eg: slightly less stable therefore more likely to form seed PrP(Sc)?)
  • Can AAV work as an antisense vector for gain of function, as well as loss of function diseases?
  • Fairly large French population exposed to contaminated growth hormone – Is anyone studying who, of those exposed, is contracting CJD?  Can we learn anything about resistance factors from tracking this situation?
  • What other mechanisms/lifestyle choices, besides exercise, to promote protein turnover/ autophagy?
  • Antibodies don’t enter cells, but amyloid formation is intracellular in FFI.  Is immunotherapy still a viable strategy?
  • Link to insomnia and development of sleep meds – how to interest researchers and/or pharmaceutical companies?
  • Better elucidate link between bone marrow, microglia, astrocytes, and disease
  • Feasibility of developing primers and sequencing PRNP gene in order to check for resistance polymorphisms (which in theory are extremely geographically specific, but this could just reflect lack of data?)  Tools: NCBI/GenBank, Ensembl, Frodo, Sigma Aldrich.  Cautions: look for secondary structures; avoid “primer dimers” (primers that anneal to each other instead of to DNA); don’t want too many C-G bonds (harder to denature); set primers around 100 base pairs out in either direction (so that noise doesn’t interfere with site of interest.)