Lentivector-mediated RNAi efficiently suppresses prion protein and prolongs survival of scrapie-infected mice.
Prion diseases are fatal neurodegenerative diseases characterized by the accumulation of PrP(Sc), the infectious and protease-resistant form of the cellular prion protein (PrP(C)). We generated lentivectors expressing PrP(C)-specific short hairpin RNAs (shRNAs) that efficiently silenced expression of the prion protein gene (Prnp) in primary neuronal cells. Treatment of scrapie-infected neuronal cells with these lentivectors resulted in an efficient and stable suppression of PrP(Sc) accumulation. After intracranial injection, lentiviral shRNA reduced PrP(C) expression in transgenic mice carrying multiple copies of Prnp. To test the therapeutic potential of lentiviral shRNA, we used what we believe to be a novel approach in which the clinical situation was mimicked. We generated chimeric mice derived from lentivector-transduced embryonic stem cells. Depending on the degree of chimerism, these animals carried the lentiviral shRNAs in a certain percentage of brain cells and expressed reduced levels of PrP(C). Importantly, in highly chimeric mice, survival after scrapie infection was significantly extended. Taken together, these data suggest that lentivector-mediated RNA interference could be an approach for the treatment of prion disease.
Pfeifer
Alexander
A
Institute of Pharmacology and Toxicology, University of Bonn, Bonn, Germany. alexander.pfeifer@uni-bonn.de
Eigenbrod
Sabina
S
Al-Khadra
Saba
S
Hofmann
Andreas
A
Mitteregger
Gerda
G
Moser
Markus
M
Bertsch
Uwe
U
Kretzschmar
Hans
H
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018621:PrPC Proteins / Q000235:genetics / Q000378:metabolism; D018620:PrPSc Proteins / Q000235:genetics / Q000378:metabolism; D017096:Prion Diseases / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D011328:Prions / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012608:Scrapie / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D012756:Sheep; D016019:Survival Analysis
D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins