A Brain Aggregate Model Gives New Insights Into the Pathobiology and Treatment of Prion Diseases

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 (PrPSc). Their abundant dendrites undergo degeneration shortly after prion infection. Treatment of prion-infected BrnAggs with drugs, such as a F-secretase inhibitors and quinacrine (Qa), which stop PrPSc formation and dendritic degeneration, mirrors the results from rodent studies. Because PrPSc 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 PrPSc 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 PrPSc by 56% compared with that of Qa alone, which reduced PrPSc by 37% relative to Rap alone. We conclude that the decrease was dominated by the ability of Qa to decrease the formation of PrPSc. Therefore, BrnAggs provide an efficient in vitro tool for screening drug therapies and studying the complex biology of prions.