title: Potential therapeutic target discovery by 2D-DIGE proteomic analysis in mouse models of asthma.
abstract: As asthma physiopathology is complex and not fully understood to date; it is expected that new key mediators are still to be unveiled in this disease. The main objective of this study was to discover potential new target proteins with a molecular weight >20 kDa by using two-dimensional differential in-gel electrophoresis (2D-DIGE) on lung parenchyma extracts from control or allergen-exposed mice (ovalbumin). Two different mouse models leading to the development of acute airway inflammation (5 days allergen exposure) and airway remodeling (10 weeks allergen exposure) were used. This experimental setting allowed the discrimination of 33 protein spots in the acute inflammation model and 31 spots in the remodeling model displaying a differential expression. Several proteins were then identified by MALDI-TOF/TOF MS. Among those differentially expressed proteins, PDIA6, GRP78, Annexin A6, hnRPA3, and Enolase display an increased expression in lung parenchyma from mice exposed to allergen for 5 days. Conversely, Apolipoprotein A1 was shown to be decreased after allergen exposure in the same model. Analysis on lung parenchyma of mice exposed to allergens for 10 weeks showed decreased calreticulin levels. Changes in the levels of those different mediators were confirmed by Western blot and immunohistochemical analysis. Interestingly, alveolar macrophages isolated from lungs in the acute inflammation model displayed enhanced levels of GRP78. Moreover, intratracheal instillation of anti-GRP78 siRNA in allergen-exposed animals led to a decrease in eosinophilic inflammation and bronchial hyperresponsiveness. This study unveils new mediators of potential importance that are up- and down-regulated in asthma. Among up-regulated mediators, GRP-78 appears as a potential new therapeutic target worthy of further investigations.
authors.affiliation: GIGA-research, GIGA-I3, GIGA-cancer, and GIGA-Neurosciences, University of Liege, Belgium.
authors.forename: Florence
authors.initials: F
authors.lastname: Quesada Calvo
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
authors.forename: Jenny
authors.initials: J
authors.lastname: Renaut
authors.forename: Céline
authors.initials: C
authors.lastname: Crahay
authors.forename: Maud
authors.initials: M
authors.lastname: Gueders
authors.forename: Jonathan
authors.initials: J
authors.lastname: Hacha
authors.forename: Geneviève
authors.initials: G
authors.lastname: Paulissen
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Foidart
authors.forename: Agnes
authors.initials: A
authors.lastname: Noel
authors.forename: Natacha
authors.initials: N
authors.lastname: Rocks
authors.forename: Pierre
authors.initials: P
authors.lastname: Leprince
authors.forename: Didier
authors.initials: D
authors.lastname: Cataldo
mesh_terms: D056151:Airway Remodeling / Q000502:physiology; D000485:Allergens / Q000276:immunology; D000818:Animals; D001249:Asthma / Q000378:metabolism*; D015153:Blotting, Western; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D055808:Drug Discovery / Q000379:methods*; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D016676:Macrophages, Alveolar / Q000378:metabolism; D008297:Male; D051379:Mice; D011014:Pneumonia / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D000485:Allergens; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D020543:Proteome