title: iRAGE as a novel carboxymethylated peptide that prevents advanced glycation end product-induced apoptosis and endoplasmic reticulum stress in vascular smooth muscle cells.
abstract: Advanced glycation end-products (AGE) and the receptor for AGE (RAGE) have been linked to numerous diabetic vascular complications. RAGE activation promotes a self-sustaining state of chronic inflammation and has been shown to induce apoptosis in various cell types. Although previous studies in vascular smooth muscle cells (VSMC) showed that RAGE activation increases vascular calcification and interferes with their contractile phenotype, little is known on the potential of RAGE to induce apoptosis in VSMC. Using a combination of apoptotic assays, we showed that RAGE stimulation with its ligand CML-HSA promotes apoptosis of VSMC. The formation of stress granules and the increase in the level of the associated protein HuR point toward RAGE-dependent endoplasmic reticulum (ER) stress, which is proposed as a key contributor of RAGE-induced apoptosis in VSMC as it has been shown to promote cell death via numerous mechanisms, including up-regulation of caspase-9. Chronic NF-κB activation and modulation of Bcl-2 homologs are also suspected to contribute to RAGE-dependent apoptosis in VSMC. With the goal of reducing RAGE signaling and its detrimental impact on VSMC, we designed a RAGE antagonist (iRAGE) derived from the primary amino acid sequence of HSA. The resulting CML peptide was selected for the high glycation frequency of the primary sequence in the native protein in vivo. Pretreatment with iRAGE blocked 69.6% of the increase in NF-κB signaling caused by RAGE activation with CML-HSA after 48h. Preincubation with iRAGE was successful in reducing RAGE-induced apoptosis, as seen through enhanced cell survival by SPR and reduced PARP cleavage. Activation of executioner caspases was 63.5% lower in cells treated with iRAGE before stimulation with CML-HSA. To our knowledge, iRAGE is the first antagonist shown to block AGE-RAGE interaction and we propose the molecule as an initial candidate for drug discovery.
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada.
authors.forename: Jean-Sébastien
authors.initials: JS
authors.lastname: Maltais
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada.
authors.forename: Elie
authors.initials: E
authors.lastname: Simard
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Froehlich
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada.
authors.forename: Jean-Bernard
authors.initials: JB
authors.lastname: Denault
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada.
authors.forename: Louis
authors.initials: L
authors.lastname: Gendron
authors.affiliation: Département de pharmacologie et physiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada. Electronic address: Michel.Grandbois@USherbrooke.ca.
authors.forename: Michel
authors.initials: M
authors.lastname: Grandbois
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002460:Cell Line; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D017127:Glycation End Products, Advanced / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D000067759:Receptor for Advanced Glycation End Products / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D017127:Glycation End Products, Advanced; D016328:NF-kappa B; D010455:Peptides; D019253:Proto-Oncogene Proteins c-bcl-2; D000067759:Receptor for Advanced Glycation End Products; D012709:Serum Albumin; C048496:N(6)-carboxymethyllysine; D053148:Caspase 3; D053179:Caspase 7; D008239:Lysine
keywords: Apoptosis; Diabetes; ER stress; Glycation; NF-κB; RAGE