A multimodal RAGE-specific inhibitor reduces amyloid β–mediated brain disorder in a mouse model of Alzheimer disease
Rashid Deane, … , Benjamin L. Miller, Berislav V. Zlokovic
Rashid Deane, … , Benjamin L. Miller, Berislav V. Zlokovic
Published March 12, 2012
Citation Information: J Clin Invest. 2012;122(4):1377-1392. https://doi.org/10.1172/JCI58642.
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Research Article Neuroscience

A multimodal RAGE-specific inhibitor reduces amyloid β–mediated brain disorder in a mouse model of Alzheimer disease

Abstract

In Alzheimer disease (AD), amyloid β peptide (Aβ) accumulates in plaques in the brain. Receptor for advanced glycation end products (RAGE) mediates Aβ-induced perturbations in cerebral vessels, neurons, and microglia in AD. Here, we identified a high-affinity RAGE-specific inhibitor (FPS-ZM1) that blocked Aβ binding to the V domain of RAGE and inhibited Aβ40- and Aβ42-induced cellular stress in RAGE-expressing cells in vitro and in the mouse brain in vivo. FPS-ZM1 was nontoxic to mice and readily crossed the blood-brain barrier (BBB). In aged APPsw/0 mice overexpressing human Aβ-precursor protein, a transgenic mouse model of AD with established Aβ pathology, FPS-ZM1 inhibited RAGE-mediated influx of circulating Aβ40 and Aβ42 into the brain. In brain, FPS-ZM1 bound exclusively to RAGE, which inhibited β-secretase activity and Aβ production and suppressed microglia activation and the neuroinflammatory response. Blockade of RAGE actions at the BBB and in the brain reduced Aβ40 and Aβ42 levels in brain markedly and normalized cognitive performance and cerebral blood flow responses in aged APPsw/0 mice. Our data suggest that FPS-ZM1 is a potent multimodal RAGE blocker that effectively controls progression of Aβ-mediated brain disorder and that it may have the potential to be a disease-modifying agent for AD.

Authors

Rashid Deane, Itender Singh, Abhay P. Sagare, Robert D. Bell, Nathan T. Ross, Barbra LaRue, Rachal Love, Sheldon Perry, Nicole Paquette, Richard J. Deane, Meenakshisundaram Thiyagarajan, Troy Zarcone, Gunter Fritz, Alan E. Friedman, Benjamin L. Miller, Berislav V. Zlokovic

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Figure 2

FPS-ZM1 and FPS2 inhibit Aβ/RAGE binding in cell-free and cell-based assays.

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FPS-ZM1 and FPS2 inhibit Aβ/RAGE binding in cell-free and cell-based ass...
(A) 125I-Aβ40 (5 nM) binding to immobilized human sRAGE in the presence of FPS2 or FPS-ZM1 (10–500 nM). (B) 125I-HMGB1 (5 nM) or 125I-S100B (5 nM) binding to sRAGE in the presence of FPS-ZM1 (10–1,000 nM). In A and B, Ki represents inhibitory constant. (C) 125I-Aβ40 (5 nM) binding to immobilized human recombinant RAGE V domain (Vd) or C1C2 domain (C1C2d) with and without FPS-ZM1 (200 nM) and to sRAGE with and without RAGE-specific C1 domain (anti-C1d), C2 domain (anti-C2d), or V domain (anti-Vd) antibodies (20 μg/ml), NI-IgG, and FPS-ZM1 (100 nM). (D) Aβ40 and Aβ42 binding to immobilized human sLRP with and without FPS-ZM1 (1 μM) or RAP (1 μM). (E) Aβ40-induced (1 μM) TBARS in RAGE-CHO cells in the presence of vehicle (closed triangle) or various concentrations of FPS2 (white circles) and FPS-ZM1 (black circles). (F) Aβ40-induced (1 μM) NF-κB activation in RAGE-CHO cells with and without FPS2 and FPS-ZM1. (G–I) BACE1 mRNA (G), BACE1 protein (H), and secreted sAPPβ (I) levels determined by qRT-PCR, immunoblotting, and ELISA, respectively, in SH-SY5Y cultures treated with vehicle or Aβ40 (1 μM) with or without FPS2 or FPS-ZM1 (50 nM) after transduction with Ad. GFP or a mutant Ad. IκB-α (S32, 36A), and/or transfection with scrambled siRNA or RAGE-siRNA. All values are means ± SEM. n = 3–5 independent experiments. β-actin was used as a loading control in H.