Journal Article

Discovery of a Novel, Monocationic, Small-Molecule Inhibitor of Scrapie Prion Accumulation in Cultured Sheep Microglia and Rov Cells

Balmer, Bethany F, Stanton, James B, Mathison, Bruce A, Schneider, David A, Kumar, Arvind, Chiesa, Roberto, Baszler, Timothy V, Dinkel, Kelcey D, Boykin, David W

2012

Overview

Prion diseases, including sheep scrapie, are neurodegenerative diseases with the fundamental pathogenesis involving conversion of normal cellular prion protein (PrP C) to disease-associated prion protein (PrP Sc). Chemical inhibition of prion accumulation is widely investigated, often using rodent-adapted prion cell culture models. Using a PrP Sc -specific ELISA we discovered a monocationic phenyl-furan-benzimidazole (DB772), which has previously demonstrated anti-pestiviral activity and represents a chemical category previously untested for anti-prion activity, that inhibited PrP Sc accumulation and prion infectivity in primary sheep microglial cell cultures (PRNP 136VV/154RR/171QQ) and Rov9 cultures (VRQ-ovinized RK13 cells). We investigated potential mechanisms of this anti-prion activity by evaluating PrP C expression with quantitative RT-PCR and PrP ELISA, comparing the concentration-dependent anti-prion and anti-pestiviral effects of DB772, and determining the selectivity index. Results demonstrate at least an approximate two-log inhibition of PrP Sc accumulation in the two cell systems and confirmed that the inhibition of PrP Sc accumulation correlates with inhibition of prion infectivity. PRNP transcripts and total PrP protein concentrations within cell lysates were not decreased; thus, decreased PrP C expression is not the mechanism of PrP Sc inhibition. PrP Sc accumulation was multiple logs more resistant than pestivirus to DB772, suggesting that the anti-PrP Sc activity was independent of anti-pestivirus activity. The anti-PrP Sc selectivity index in cell culture was approximately 4.6 in microglia and 5.5 in Rov9 cells. The results describe a new chemical category that inhibits ovine PrP Sc accumulation in primary sheep microglia and Rov9 cells, and can be used for future studies into the treatment and mechanism of prion diseases.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Accumulation / Analysis / Animal models / Animals / Antimicrobial agents / Benzimidazole / Benzimidazoles / Benzimidazoles - chemistry / Benzimidazoles - pharmacology / Biology / Cations / Cell culture / Cell Death - drug effects / Cells, Cultured / Chemistry / Chemotherapy / Creutzfeldt-Jakob disease / Curcumin - pharmacology / Diarrhea / Diseases / Enzyme-linked immunosorbent assay / Fibroblasts / Furans - chemistry / Furans - pharmacology / Infections / Infectivity / Inhibition / Lysates / Medical treatment / Medicine / Microglia / Microglia - drug effects / Microglia - metabolism / Nervous system diseases / Neurodegenerative diseases / neuroglia / Neurological diseases / Odocoileus / Ovis aries / Pathogenesis / pathogenicity / Pathology / Pestivirus / Pestivirus - drug effects / Polyamines / Polymerase chain reaction / Prion protein / Prions / Prions (Proteins) / Prions - genetics / Prions - metabolism / Proteins / PrPSc proteins / PrPSc Proteins - antagonists & inhibitors / PrPSc Proteins - pathogenicity / reverse transcriptase polymerase chain reaction / RNA, Messenger - genetics / RNA, Messenger - metabolism / Scrapie / Scrapie - metabolism / Scrapie - pathology / Selectivity / Sheep / Small Molecule Libraries - chemistry / Small Molecule Libraries - pharmacology / Studies / Thiophene / Veterinary Science / Viruses