Site-Specific Blockade of RAGE-Vd Prevents Amyloid- Oligomer Neurotoxicity

Department of Pediatrics, Division of Clinical Chemistry and Biochemistry, University of Zurich, 8032 Zurich, Switzerland.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience (Impact Factor: 6.34). 06/2008; 28(20):5149-58. DOI: 10.1523/JNEUROSCI.4878-07.2008
Source: PubMed


In the genesis of Alzheimer's disease (AD), converging lines of evidence suggest that amyloid-beta peptide (Abeta) triggers a pathogenic cascade leading to neuronal loss. It was long assumed that Abeta had to be assembled into extracellular amyloid fibrils or aggregates to exert its cytotoxic effects. Over the past decade, characterization of soluble oligomeric Abeta species in the brains of AD patients and in transgenic models has raised the possibility that different conformations of Abeta may contribute to AD pathology via different mechanisms. The receptor for advanced glycation end products (RAGE), a member of the Ig superfamily, is a cellular binding site for Abeta. Here, we investigate the role of RAGE in apoptosis induced by distinct well characterized Abeta conformations: Abeta oligomers (AbetaOs), Abeta fibrils (AbetaFs), and Abeta aggregates (AbetaAs). In our in vitro system, treatment with polyclonal anti-RAGE antibodies significantly improves SHSY-5Y cell and neuronal survival exposed to either AbetaOs or AbetaAs but does not affect AbetaF toxicity. Interestingly, using site-specific antibodies, we demonstrate that targeting of the V(d) domain of RAGE attenuates AbetaO-induced toxicity in both SHSY-5Y cells and rat cortical neurons, whereas inhibition of AbetaA-induced apoptosis requires the neutralization of the C(1d) domain of the receptor. Thus, our data indicate that distinct regions of RAGE are involved in Abeta-induced cellular and neuronal toxicity with respect to the Abeta aggregation state, and they suggest the blockage of particular sites of the receptor as a potential therapeutic strategy to attenuate neuronal death.

16 Reads
  • Source
    • "As reported in the literature, the receptor for advanced glycation end products (RAGE) plays an important role as a cell-surface receptor for Aβ at the blood-brain barrier and in neurons and microglia (Yan et al. 1996, 2010; Zlokovic 2011; Origlia et al. 2010). The extracellular V domain of RAGE is the key domain in Aβ binding to RAGE (Sturchler et al. 2008). In the Aβ peptide, the major binding site is localized to an eight amino acid stretch of residues in positions 16–23 of the KLVFFAED sequence, which consists of a series of hydrophobic residues flanked by two negatively charged residues at the peptide c-terminus (Gospodarska et al. 2011; Jargilo et al. 2013). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Alzheimer's disease (AD) is an age-related neurodegenerative disorder in which amyloid β (Aβ) peptide accumulates in the brain. The receptor for advanced glycation end product (RAGE) is a cellular binding site for Aβ peptide and mediates amyloid β-induced perturbations in cerebral vessels, neurons, and microglia in AD. Here, we identified a specific high-affinity RAGE inhibitor (APDTKTQ named RP-1) from a phage display library. RP-1 bound to RAGE and inhibited Aβ peptide-induced cellular stress in human neuroblastoma SH-SYSY cells in vitro. Three amino acids in RP-1 are identical to those in the Aβ peptide. RP-1 shows high homology to the 16-23 (KLVFFAED) regions in Aβ peptide and high-affinity RAGE. Functional analyses indicated that RP-1 significantly reduced the level of reactive oxygen species (ROS) and ROS products and that it enhanced catalase and glutathione peroxidase (GPx) activity. Furthermore, it inactivated caspase3 and caspase9 and inhibited the upregulation of RAGE, nuclear factor-κB (NF-κB), and beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1) protein expression. In addition, RP-1 activated the PI3K/AKT signaling pathway, inhibiting the interaction between Bax and Bcl-2. Our data suggest that RP-1 is a potent RAGE blocker that effectively controls the progression of Aβ peptide-mediated brain disorders and that it may have potential as a disease-modifying agent for AD.
    Applied Microbiology and Biotechnology 10/2015; DOI:10.1007/s00253-015-7001-7 · 3.34 Impact Factor
  • Source
    • "When indicated , a polyclonal anti-S100B antibody (DakoCytomation) or polyclonal anti-RAGE antibody (100 nM) was added to the cells at the time point of seeding into wells. The polyclonal anti-RAGE antibody consisted of an equimolar mixture of anti-V, anti-C1 and anti-C2 antibodies as described in our previous publications [34] [35]. 2.7.3. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The formation of melanoma metastases from primary tumor cells is a complex phenomenon that involves the regulation of multiple genes. We have previously shown that the receptor for advanced glycation end products (RAGE) was up-regulated in late metastatic stages of melanoma patient samples and we hypothesized that up-regulation of RAGE in cells forming a primary melanoma tumor could contribute to the metastatic switch of these cells. To test our hypothesis, we overexpressed RAGE in the WM115 human melanoma cell line that was established from a primary melanoma tumor of a patient. We show here that overexpression of RAGE in these cells is associated with mesenchymal-like morphologies of the cells. These cells demonstrate higher migration abilities and reduced proliferation properties, suggesting that the cells have switched to a metastatic phenotype. At the molecular level, we show that RAGE overexpression is associated with the up-regulation of the RAGE ligand S100B and the down-regulation of p53, ERK1/2, cyclin E and NF-kB. Our study supports a role of RAGE in the metastatic switch of melanoma cells.
    Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease 07/2014; 1842(7). DOI:10.1016/j.bbadis.2014.02.013 · 4.88 Impact Factor
  • Source
    • "Also, due to the presence of NF-κB transcriptional elements in its gene promoter [19,20], RAGE activation causes its own sustained over-expression, converting a transient proinflammatory response into a chronic proinflammatory signal [21]. Such positive feedback loop is a potential therapeutic target, and RAGE has been suggested as a biomarker and/or target for intervention in several diseases, including diabetes and its complications [22,23], inflammation [21], tumors [18] and neurodegeneration [2]. Rational design of therapeutic RAGE antagonists or modulators of RAGE activity requires sound structural knowledge of the receptor itself, its structural forms, and its complexes with ligands. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Activation of the receptor for advanced glycation end products (RAGE) leads to a chronic proinflammatory signal, affecting patients with a variety of diseases. Potentially beneficial modification of RAGE activity requires understanding the signal transduction mechanism at the molecular level. The ligand binding domain is structurally uncoupled from the cytoplasmic domain, suggesting receptor oligomerization is a requirement for receptor activation. In this study, we used hydrogen-deuterium exchange and mass spectrometry to map structural differences between the monomeric and oligomeric forms of RAGE. Our results indicated the presence of a region shielded from exchange in the oligomeric form of RAGE and led to the identification of a new oligomerization interface localized at the linker region between domains C1 and C2. Based on this finding, a model of a RAGE dimer and higher oligomeric state was constructed.
    PLoS ONE 10/2013; 8(10):e76353. DOI:10.1371/journal.pone.0076353 · 3.23 Impact Factor
Show more


16 Reads
Available from