Gene expression in whole lung and pulmonary macrophages reflects the dynamic pathology associated with airway surface dehydration

Abstract

BackgroundDefects in airway mucosal defense, including decreased mucus clearance, contribute to the pathogenesis of human chronic obstructive pulmonary diseases. Scnn1b-Tg mice, which exhibit chronic airway surface dehydration from birth, can be used as a model to study the pathogenesis of muco-obstructive lung disease across developmental stages. To identify molecular signatures associated with obstructive lung disease in this model, gene expression analyses were performed on whole lung and purified lung macrophages collected from Scnn1b-Tg and wild-type (WT) littermates at four pathologically relevant time points. Macrophage gene expression at 6 weeks was evaluated in mice from a germ-free environment to understand the contribution of microbes to disease development.ResultsDevelopment- and disease-specific shifts in gene expression related to Scnn1b over-expression were revealed in longitudinal analyses. While the total number of transgene-related differentially expressed genes producing robust signals was relatively small in whole lung (n = 84), Gene Set Enrichment Analysis (GSEA) revealed significantly perturbed biological pathways and interactions between normal lung development and disease initiation/progression. Purified lung macrophages from Scnn1b-Tg mice exhibited numerous robust and dynamic gene expression changes. The expression levels of Classically-activated (M1) macrophage signatures were significantly altered at post-natal day (PND) 3 when Scnn1b-Tg mice lung exhibit spontaneous bacterial infections, while alternatively-activated (M2) macrophage signatures were more prominent by PND 42, producing a mixed M1-M2 activation profile. While differentially-regulated, inflammation-related genes were consistently identified in both tissues in Scnn1b-Tg mice, there was little overlap between tissues or across time, highlighting time- and tissue-specific responses. Macrophages purified from adult germ-free Scnn1b-Tg mice exhibited signatures remarkably similar to non-germ-free counterparts, indicating that the late-phase macrophage activation profile was not microbe-dependent.ConclusionsWhole lung and pulmonary macrophages respond independently and dynamically to local stresses associated with airway mucus stasis. Disease-specific responses interact with normal developmental processes, influencing the final state of disease in this model. The robust signatures observed in Scnn1b-Tg lung macrophages highlight their critical role in disease pathogenesis. These studies emphasize the importance of region-, cell-type-, and time-dependent analyses to fully dissect the natural history of disease and the consequences of disease on normal lung development.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-726) contains supplementary material, which is available to authorized users.

UDP-glucose promotes neutrophil recruitment in the lung

“…5) could reflect a contribution of P2Y 14 R-promoted cytokine release from alveolar epithelial cells, upstream of neutrophil recruitment [47]. In addition to the above-discussed scenario, published gene expression analyses indicated high levels of P2Y 14 R mRNA in murine alveolar macrophages [48]. Therefore, activation of P2Y 14 R on lung resident macrophages also potentially results in production of pro-inflammatory cytokines leading to neutrophil recruitment.…”

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“…Evaluation of muco-obstructive disease development in the presence and absence of MFs is one way to investigate the complex interrelationships between mucus stasis and MF function. This approach is challenging, however, as the pathophysiology associated with both MF depletion and mucostasis are superimposed on normal lung developmental processes in both of our models (9). Furthermore, MF activation is known to be highly pleiotropic and sensitive to local environmental stimuli (17).…”

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