title: A brain aggregate model gives new insights into the pathobiology and treatment of prion diseases.
abstract: Brain aggregates (BrnAggs) derived from fetal mouse brains contain mature neurons and glial cells. We determined that BrnAggs are consistently infected with Rocky Mountain Laboratory scrapie strain prions and produce increasing levels of the pathogenic form of the prion protein (PrP). Their abundant dendrites undergo degeneration shortly after prion infection. Treatment of prion-infected BrnAggs with drugs, such as a γ-secretase inhibitors and quinacrine (Qa), which stop PrP formation and dendritic degeneration, mirrors the results from rodent studies. Because PrP is trafficked into lysosomes by endocytosis and autophagosomes by phagocytosis in neurons of prion strain-infected BrnAggs, we studied the effects of drugs that modulate subcellular trafficking. Rapamycin (Rap), which activates autophagy, markedly increased light-chain 3-II (LC3-II)-positive autophagosomes and cathepsin D-positive lysosomes in BrnAggs but could not eliminate the intracellular PrP within them. Adding Qa to Rap markedly reduced the number of LC3-II-positive autolysosomes. Rap + Qa created a competition between Rap increasing and Qa decreasing LC3-II. Rapamycin + Qa decreased total PrP by 56% compared with that of Qa alone, which reduced PrP by 37% relative to Rap alone. We conclude that the decrease was dominated by the ability of Qa to decrease the formation of PrP. Therefore, BrnAggs provide an efficient in vitro tool for screening drug therapies and studying the complex biology of prions.
authors.affiliation: Department of Pathology, University of California, San Francisco, USA.
authors.forename: Krystyna
authors.initials: K
authors.lastname: Bajsarowicz
authors.forename: Misol
authors.initials: M
authors.lastname: Ahn
authors.forename: Larry
authors.initials: L
authors.lastname: Ackerman
authors.forename: Bernadette N
authors.initials: BN
authors.lastname: Dearmond
authors.forename: George
authors.initials: G
authors.lastname: Carlson
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: DeArmond
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001381:Azepines / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000196:embryology / Q000378:metabolism* / Q000473:pathology*; D002402:Cathepsin D / Q000378:metabolism; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D007166:Immunosuppressive Agents / Q000494:pharmacology; D066298:In Vitro Techniques; D008247:Lysosomes / Q000502:physiology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D017628:Microglia / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009410:Nerve Degeneration / Q000209:etiology / Q000473:pathology / Q000517:prevention & control; D009419:Nerve Tissue Proteins / Q000378:metabolism; D018620:PrPSc Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011247:Pregnancy; D017096:Prion Diseases / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D000072002:Prion Proteins; D011328:Prions / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D011796:Quinacrine / Q000494:pharmacology; D020123:Sirolimus / Q000494:pharmacology; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013997:Time Factors
chemical_list: D001381:Azepines; D004791:Enzyme Inhibitors; D007166:Immunosuppressive Agents; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; C484278:N2-((2S)-2-(3,5-difluorophenyl)-2-hydroxyethanoyl)-N1-((7S)-5-methyl-6-oxo-6,7-dihydro-5H-dibenzo(b,d)azepin-7-yl)-L-alaninamide; D009419:Nerve Tissue Proteins; D018620:PrPSc Proteins; D000072002:Prion Proteins; D011328:Prions; C512657:Prnp protein, mouse; D002402:Cathepsin D; D011796:Quinacrine; D000409:Alanine; D020123:Sirolimus