Abstract
A key feature of prion diseases is the conversion of the cellular prion protein (PrP(C)) into disease-related isoforms (PrP(Sc)), the deposition of which is thought to lead to neurodegeneration. In this study a pharmacological approach was used to determine the metabolic pathways involved in the formation of protease-resistant PrP (PrP(res)) in three prion-infected cell lines (ScN2a, SMB, and ScGT1 cells). Daily treatment of these cells with phospholipase A(2) (PLA(2)) inhibitors for 7 days prevented the accumulation of PrP(res). Glucocorticoids with anti-PLA(2) activity also prevented the formation of PrP(res) and reduced the infectivity of SMB cells. Treatment with platelet-activating factor (PAF) antagonists also reduced the PrP(res) content of cells, while the addition of PAF reversed the inhibitory effect of PLA(2) inhibitors on PrP(res) formation. ScGT1 cells treated with PLA(2) inhibitors or PAF antagonists for 7 days remained clear of detectable (PrPres) when grown in control medium for a further 12 weeks. Treatment of non-infected cells with PLA(2) inhibitors or PAF antagonists reduced PrP(C) levels suggesting that limiting cellular PrP(C) may restrict prion formation in infected cells. These data indicate a pivotal role for PLA(2) and PAF in controlling PrP(res) formation and identify them as potential therapeutic agents.
Highlights
A key feature of prion diseases is the conversion of the cellular prion protein (PrPC) into disease-related isoforms (PrPSc), the deposition of which is thought to lead to neurodegeneration
phospholipase A2 (PLA2) Inhibitors Reduce the PrPres Content of Three Prioninfected Cell Lines—In an initial screening experiment, the effects of drugs that inhibit some of the common signal transduction pathways were investigated for their effects on the PrPres content of ScN2a cells
In the present study we utilized a pharmacological approach to determine the metabolic pathways that underlie the formation of PrPres in three prion-infected neuroblastoma cell lines (ScN2a, ScGT1, and SMB cells)
Summary
University College, Dublin, Ireland) that produce PrPSc and infectious agent, were grown in Hams F12 medium containing 2 mM glutamine, standard antibiotics (100 units/ml penicillin and 100 g/ml streptomycin) and 2% fetal calf serum. SMB cells (TSE Resource Centre, Institute for Animal Health, Compton, UK), which produce PrPSc and infectious agent, were grown in RPMI 1640 medium containing standard antibiotics, 2 mM glutamine and 2% fetal calf serum. ScGT1 cells (supplied by Dr Sylvain Lehmann, CNRSIGH, Montpellier, France), an immortalized murine hypothalamic neuronal cell line infected by the scrapie Chandler isolate and that persistently expresses PrPres, were grown in Optimem supplemented with 2 mM glutamine, 5% fetal calf serum, and standard antibiotics. Cells were grown with daily changes of media and PrPres production was evaluated after 7 days.
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