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Potential therapeutic target discovery by 2D-DIGE proteomic analysis in mouse models of asthma. | LitMetric

AI Article Synopsis

  • The study investigates the complex nature of asthma and aims to identify new target proteins related to the disease using two-dimensional differential in-gel electrophoresis (2D-DIGE) on lung tissues from mice exposed to allergens.
  • Two mouse models were used: one for acute airway inflammation (5 days of allergen exposure) and another for airway remodeling (10 weeks of exposure), revealing 33 and 31 differentially expressed protein spots, respectively.
  • Notable proteins identified include GRP78, which was found to be up-regulated in acute inflammation and linked to reduced inflammation when targeted with anti-GRP78 siRNA, suggesting it could be a promising new therapeutic target for asthma.

Article 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.

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Source
http://dx.doi.org/10.1021/pr200494nDOI Listing

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