Dual MicroRNA to Cellular Prion Protein Inhibits Propagation of Pathogenic Prion Protein in Cultured Cells

Abstract

Prion diseases are fatal transmissible neurodegenerative disorders affecting humans and various mammals. In spite of intensive efforts, there is no effective cure or treatment for prion diseases. Cellular forms of prion protein (PrP^C) is essential for propagation of abnormal isoforms of prion protein (PrP^Sc) and pathogenesis. The effect of an artificial dual microRNA (DmiR) on PrP^C suppression and resultant inhibition of prion replication was determined using prion-infectible cell cultures: differentiated C2C12 culture and primary mixed neuronal and glial cells culture (MNGC). Processing of DmiR by prion-susceptible myotubes, but not by reserve cells, in differentiated C2C12 culture slowed prion replication, implying an importance of cell type-specific PrP^C targeting. In MNGC, reduction of PrP^C with DmiR was effective for suppressing prion replication. MNGC lentivirally transduced with non-targeting control miRNAs (scrambled) reduced prion replication at a level similar to that with a synthetic analogue of viral RNA, poly I:C. The results suggest that a synergistic combination of the immunostimulatory RNA duplexes (miRNA) and PrP^C silencing with DmiR might augment a therapeutic potential of RNA interference.

Keywords: Differentiated C2C12 cells; Immune stimulator; Neuronal and glial cells; Prion disease; Therapeutic RNAi.