Related Experiment Video
Updated: Jul 13, 2026
Enrichment of Mammalian Tissues and Xenopus Oocytes with Cholesterol
Published on: March 25, 2020
Antiprion activity of cholesterol esterification modulators: a comparative study using ex vivo sheep fibroblasts and
Alessandra Pani1, Claudia Norfo, Claudia Abete
1Department of Biomedical Sciences and Technologies, University of Cagliari, 09042-Monserrato, Italy. pania@unica.it
Abstract:
Our studies on the role of cholesterol homeostasis in the pathogenesis of scrapie revealed abnormal accumulation of cholesterol esters in ex vivo peripheral blood mononuclear cells (PBMCs) and skin fibroblasts from healthy and scrapie-affected sheep carrying a scrapie-susceptible genotype compared to sheep with a resistant genotype. Similar alterations were observed in mouse neuroblastoma N2a cell lines persistently infected with mouse-adapted 22L and RML strains of scrapie that showed up to threefold-higher cholesterol ester levels than parental N2a cells. We now report that proteinase K-resistant prion protein (PrPres)-producing cell populations of subclones from scrapie-infected cell lines were characterized by higher cholesterol ester levels than clone populations not producing PrPres. Treatments with a number of drugs known to interfere with different steps of cholesterol metabolism strongly reduced the accumulation of cholesterol esters in ex vivo PBMCs and skin fibroblasts from scrapie-affected sheep but had significantly less or no effect in their respective scrapie-resistant or uninfected counterparts. In scrapie-infected N2a cells, inhibition of cholesterol esters was associated with selective antiprion activity. Effective antiprion concentrations of cholesterol modulators (50% effective concentration [EC(50)] range, 1.4 to 40 microM) were comparable to those of antiprion reference compounds (EC(50) range, 0.6 to 10 microM). These data confirm our hypothesis that abnormal accumulation of cholesterol esters may represent a biological marker of susceptibility to prion infection/replication and a novel molecular target of potential clinical importance.
Insights
Abnormal cholesterol ester accumulation in cells indicates susceptibility to prion diseases like scrapie. Modulating cholesterol metabolism shows potential as a therapeutic strategy against prion replication.
Area of Science:
- Neuroscience
- Biochemistry
- Pathology
Background:
- Scrapie pathogenesis involves disruptions in cellular cholesterol homeostasis.
- Previous studies indicated abnormal cholesterol ester accumulation in scrapie-affected sheep cells.
Purpose of the Study:
- To investigate the role of cholesterol ester accumulation in prion disease susceptibility and replication.
- To evaluate cholesterol modulators as a potential therapeutic target for prion diseases.
Main Methods:
- Analysis of cholesterol ester levels in peripheral blood mononuclear cells and skin fibroblasts from sheep with different scrapie genotypes.
- Assessment of cholesterol ester levels in scrapie-infected and uninfected mouse neuroblastoma N2a cell lines.
- Treatment of cells and tissues with cholesterol-modulating drugs and evaluation of antiprion activity.
Main Results:
- Scrapie-susceptible sheep cells and scrapie-infected N2a cells exhibited higher cholesterol ester levels.
- Cholesterol-modulating drugs reduced cholesterol ester accumulation in scrapie-affected cells.
- Inhibition of cholesterol esters in infected cells demonstrated selective antiprion activity.
Conclusions:
- Abnormal cholesterol ester accumulation serves as a potential biomarker for prion infection susceptibility.
- Targeting cholesterol metabolism presents a novel therapeutic avenue for prion diseases.
