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Profiling of differentially expressed genes using suppression subtractive hybridization in an equine model of chronic asthma

Lavoie Jean-Pierre, Lefebvre-Lavoie Josiane, Leclere Mathilde, Lavoie-Lamoureux Anouk, Chamberland Annie, Laprise Catherine et Lussier Jacques. (2012). Profiling of differentially expressed genes using suppression subtractive hybridization in an equine model of chronic asthma. PLoS ONE, 7, (1), e29440.

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URL officielle: http://dx.doi.org/doi:10.1371/journal.pone.0029440

Résumé

Background : Gene expression analyses are used to investigate signaling pathways involved in diseases. In asthma, they have been primarily derived from the analysis of bronchial biopsies harvested from mild to moderate asthmatic subjects and controls. Due to ethical considerations, there is currently limited information on the transcriptome profile of the peripheral lung tissues in asthma.

Objective : To identify genes contributing to chronic inflammation and remodeling in the peripheral lung tissue of horses with heaves, a naturally occurring asthma-like condition.

Methods : Eleven adult horses (6 heaves-affected and 5 controls) were studied while horses with heaves were in clinical remission (Pasture), and during disease exacerbation induced by a 30-day natural antigen challenge during stabling (Challenge). Large peripheral lung biopsies were obtained by thoracoscopy at both time points. Using suppression subtractive hybridization (SSH), lung cDNAs of controls (Pasture and Challenge) and asymptomatic heaves-affected horses (Pasture) were subtracted from cDNAs of horses with heaves in clinical exacerbation (Challenge). The differential expression of selected genes of interest was confirmed using quantitative PCR assay.

Results : Horses with heaves, but not controls, developed airway obstruction when challenged. Nine hundred and fifty cDNA clones isolated from the subtracted library were screened by dot blot array and 224 of those showing the most marked expression differences were sequenced. The gene expression pattern was confirmed by quantitative PCR in 15 of 22 selected genes. Novel genes and genes with an already defined function in asthma were identified in the subtracted cDNA library. Genes of particular interest associated with asthmatic airway inflammation and remodeling included those related to PPP3CB/NFAT, RhoA, and LTB4/GPR44 signaling pathways.

Conclusions : Pathways representing new possible targets for anti-inflammatory and anti-remodeling therapies for asthma were identified. The findings of genes previously associated with asthma validate this equine model for gene expression studies.

Type de document:Article publié dans une revue avec comité d'évaluation
Volume:7
Numéro:1
Pages:e29440
Version évaluée par les pairs:Oui
Date:2012
Sujets:Sciences de la santé > Sciences médicales > Biologie moléculaire
Sciences de la santé > Sciences médicales > Génétique
Département, module, service et unité de recherche:Départements et modules > Département des sciences fondamentales
Mots-clés:horse, asthma, gene expression profiling, polymerase chain reaction, cheval, asthme, profilage d'expression génétique, amplification par la polymérase
Informations complémentaires:This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Déposé le:28 août 2015 17:55
Dernière modification:05 déc. 2016 21:07
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