Preventing P-Gp Ubiquitination Lowers A Beta Brain Levels In An Alzheimer'S Disease Mouse Model

One characteristic of Alzheimer's disease (AD) is excessive accumulation of amyloid-beta (A beta) in the brain. A beta brain accumulation is, in part, due to a reduction in A beta clearance from the brain across the blood-brain barrier. One key element that contributes to A beta brain clearance is P-glycoprotein (P-gp) that transports A beta from brain to blood. In AD, P-gp protein expression and transport activity levels are significantly reduced, which impairs A beta brain clearance. The mechanism responsible for reduced P-gp expression and activity levels is poorly understood. We recently demonstrated that A beta(40) triggers P-gp degradation through the ubiquitin-proteasome pathway. Consistent with these data, we show here that ubiquitinated P-gp levels in brain capillaries isolated from brain samples of AD patients are increased compared to capillaries isolated from brain tissue of cognitive normal individuals. We extended this line of research to in vivo studies using transgenic human amyloid precursor protein (hAPP)-overexpressing mice (Tg2576) that were treated with PYR41, a cell-permeable, irreversible inhibitor of the ubiquitin-activating enzyme E1. Our data show that inhibiting P-gp ubiquitination protects the transporter from degradation, and immunoprecipitation experiments confirmed that PYR41 prevented P-gp ubiquitination. We further found that PYR41 treatment prevented reduction of P-gp protein expression and transport activity levels and substantially lowered A beta brain levels in hAPP mice. Together, our findings provide in vivo proof that the ubiquitin-proteasome system mediates reduction of blood-brain barrier P-gp in AD and that inhibiting P-gp ubiquitination prevents P-gp degradation and lowers A beta brain levels. Thus, targeting the ubiquitin-proteasome system may provide a novel therapeutic approach to protect blood-brain barrier P-gp from degradation in A beta and other A beta-based pathologies.