Year: 2026
Type: preprint
Abstract: ABSTRACT The normal cellular prion protein (PrP C ) is an essential substrate in all forms of prion diseases and a receptor for Aβ oligomers in Alzheimer’s disease. However, it is not fully understood how cells regulate PrP C levels. Recently, we identified glycogen synthase kinase-3β (GSK-3β) as a potential regulator of PrP C levels in a whole genome knockout screen. Here, we show that both cell surface and total PrP C levels can be reduced either by siRNA-mediated Gsk3b (but not Gsk3a ) knockdown or by CRISPR-mediated Gs3b knockout. Whole cell mass spectrometric analysis showed that PrP C was the 60 th most significantly reduced protein (out of 7227 total proteins detected) in Gsk3b knockout cells, compared to wild-type cells. Two different GSK-3 inhibitors, laduviglusib (CHIR-99021) and AZD-1080, reduced PrP C levels in mouse CAD5 and human BE(2)-C cells, both in undifferentiated and differentiated states. PrP C levels were similarly reduced by cycloheximide treatment in both Gsk3b knockout and WT cells, indicating that GSK-3β regulates PrP C levels through a post-translational mechanism. Finally, treatment with either laduviglusib or AZD-1080 reduced PrP Sc levels in CAD5 cells infected with three different rodent prion strains. Overall, the results reveal that GSK-3β activity controls PrP C levels in living cells, revealing a novel regulatory mechanism and promising therapeutic target.
Institutions: Dartmouth College, Dartmouth Cancer Center, University of Toronto
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7204239558
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