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Systems approaches to modeling chronic mucosal inflammation.

Kalita M, Tian B, Gao B, Choudhary S, Wood TG, Carmical JR, Boldogh I, Mitra S, Minna JD, Brasier AR - Biomed Res Int (2013)

Bottom Line: EMT had dramatic effects on the induction of the innate pathway and the coupling interval of the canonical and noncanonical NF- κ B pathways.Simulation experiments demonstrate that rapid, coordinated cap-independent translation of TRAF-1 and NF- κ B2 is required to reduce the noncanonical pathway coupling interval.Further applications of systems approaches will provide understanding of this complex phenotype through deterministic modeling and multidimensional (genomic and proteomic) profiling.

View Article: PubMed Central - PubMed

Affiliation: Sealy Center for Molecular Medicine, The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA.

ABSTRACT
The respiratory mucosa is a major coordinator of the inflammatory response in chronic airway diseases, including asthma and chronic obstructive pulmonary disease (COPD). Signals produced by the chronic inflammatory process induce epithelial mesenchymal transition (EMT) that dramatically alters the epithelial cell phenotype. The effects of EMT on epigenetic reprogramming and the activation of transcriptional networks are known, its effects on the innate inflammatory response are underexplored. We used a multiplex gene expression profiling platform to investigate the perturbations of the innate pathways induced by TGF β in a primary airway epithelial cell model of EMT. EMT had dramatic effects on the induction of the innate pathway and the coupling interval of the canonical and noncanonical NF- κ B pathways. Simulation experiments demonstrate that rapid, coordinated cap-independent translation of TRAF-1 and NF- κ B2 is required to reduce the noncanonical pathway coupling interval. Experiments using amantadine confirmed the prediction that TRAF-1 and NF- κ B2/p100 production is mediated by an IRES-dependent mechanism. These data indicate that the epigenetic changes produced by EMT induce dynamic state changes of the innate signaling pathway. Further applications of systems approaches will provide understanding of this complex phenotype through deterministic modeling and multidimensional (genomic and proteomic) profiling.

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TGFβ induces EMT transition in HSAECs. (a) F-actin staining. Shown in green is a confocal microscopic image of FITC-phalloidin staining merged with DAPI staining (blue) of HSAECs, A549 cells, and HBECs in the absence or presence of TGFβ stimulation as indicated. (b) Expression of EMT program. Shown is normalized mRNA expression in HSAECs in the presence or absence of EMT after 10 d of TGFβ treatment. For each gene, mRNA expression was normalized to β-pol as a housekeeping gene and expressed as the fold change relative to its expression in the absence of TGFβ stimulation. Shown is mean and SE of replicate measurements. Each point is the mean of a duplicate biological experiment, measured with three technical replicates. Abbreviations: SLUG, Snail 2; SNAL1, Snail 1; col1A, collagen type 1a.
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fig1: TGFβ induces EMT transition in HSAECs. (a) F-actin staining. Shown in green is a confocal microscopic image of FITC-phalloidin staining merged with DAPI staining (blue) of HSAECs, A549 cells, and HBECs in the absence or presence of TGFβ stimulation as indicated. (b) Expression of EMT program. Shown is normalized mRNA expression in HSAECs in the presence or absence of EMT after 10 d of TGFβ treatment. For each gene, mRNA expression was normalized to β-pol as a housekeeping gene and expressed as the fold change relative to its expression in the absence of TGFβ stimulation. Shown is mean and SE of replicate measurements. Each point is the mean of a duplicate biological experiment, measured with three technical replicates. Abbreviations: SLUG, Snail 2; SNAL1, Snail 1; col1A, collagen type 1a.

Mentions: To characterize type II EMT, HSAECs were incubated in the absence or presence of TGFβ (5 ng/mL) for 10 d. Transformed type II alveolar epithelial cells (A549) and immortalized bronchial epithelial cells (HBECs) were used as reference. Cells were fixed, stained with FITC-conjugated phalloidin (for distribution of F actin) and DAPI (a nuclear DNA stain), and examined by confocal microscopy. In the absence of TGFβ stimulation, HSAECs assumed a normal cuboidal morphology with perinuclear cytoplasmic distribution of F-actin (Figure 1(a)). By contrast, TGFβ-treated HSAECs showed an elongated shape with markedly induced F-actin staining (Figure 1(a)). This morphological change of enhanced front-rear polarity and cytoskeletal actin rearrangement are similar to those observed in TGFβ-treated A549 and HBECs; all are characteristic morphological changes of EMT [8].


Systems approaches to modeling chronic mucosal inflammation.

Kalita M, Tian B, Gao B, Choudhary S, Wood TG, Carmical JR, Boldogh I, Mitra S, Minna JD, Brasier AR - Biomed Res Int (2013)

TGFβ induces EMT transition in HSAECs. (a) F-actin staining. Shown in green is a confocal microscopic image of FITC-phalloidin staining merged with DAPI staining (blue) of HSAECs, A549 cells, and HBECs in the absence or presence of TGFβ stimulation as indicated. (b) Expression of EMT program. Shown is normalized mRNA expression in HSAECs in the presence or absence of EMT after 10 d of TGFβ treatment. For each gene, mRNA expression was normalized to β-pol as a housekeeping gene and expressed as the fold change relative to its expression in the absence of TGFβ stimulation. Shown is mean and SE of replicate measurements. Each point is the mean of a duplicate biological experiment, measured with three technical replicates. Abbreviations: SLUG, Snail 2; SNAL1, Snail 1; col1A, collagen type 1a.
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fig1: TGFβ induces EMT transition in HSAECs. (a) F-actin staining. Shown in green is a confocal microscopic image of FITC-phalloidin staining merged with DAPI staining (blue) of HSAECs, A549 cells, and HBECs in the absence or presence of TGFβ stimulation as indicated. (b) Expression of EMT program. Shown is normalized mRNA expression in HSAECs in the presence or absence of EMT after 10 d of TGFβ treatment. For each gene, mRNA expression was normalized to β-pol as a housekeeping gene and expressed as the fold change relative to its expression in the absence of TGFβ stimulation. Shown is mean and SE of replicate measurements. Each point is the mean of a duplicate biological experiment, measured with three technical replicates. Abbreviations: SLUG, Snail 2; SNAL1, Snail 1; col1A, collagen type 1a.
Mentions: To characterize type II EMT, HSAECs were incubated in the absence or presence of TGFβ (5 ng/mL) for 10 d. Transformed type II alveolar epithelial cells (A549) and immortalized bronchial epithelial cells (HBECs) were used as reference. Cells were fixed, stained with FITC-conjugated phalloidin (for distribution of F actin) and DAPI (a nuclear DNA stain), and examined by confocal microscopy. In the absence of TGFβ stimulation, HSAECs assumed a normal cuboidal morphology with perinuclear cytoplasmic distribution of F-actin (Figure 1(a)). By contrast, TGFβ-treated HSAECs showed an elongated shape with markedly induced F-actin staining (Figure 1(a)). This morphological change of enhanced front-rear polarity and cytoskeletal actin rearrangement are similar to those observed in TGFβ-treated A549 and HBECs; all are characteristic morphological changes of EMT [8].

Bottom Line: EMT had dramatic effects on the induction of the innate pathway and the coupling interval of the canonical and noncanonical NF- κ B pathways.Simulation experiments demonstrate that rapid, coordinated cap-independent translation of TRAF-1 and NF- κ B2 is required to reduce the noncanonical pathway coupling interval.Further applications of systems approaches will provide understanding of this complex phenotype through deterministic modeling and multidimensional (genomic and proteomic) profiling.

View Article: PubMed Central - PubMed

Affiliation: Sealy Center for Molecular Medicine, The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA.

ABSTRACT
The respiratory mucosa is a major coordinator of the inflammatory response in chronic airway diseases, including asthma and chronic obstructive pulmonary disease (COPD). Signals produced by the chronic inflammatory process induce epithelial mesenchymal transition (EMT) that dramatically alters the epithelial cell phenotype. The effects of EMT on epigenetic reprogramming and the activation of transcriptional networks are known, its effects on the innate inflammatory response are underexplored. We used a multiplex gene expression profiling platform to investigate the perturbations of the innate pathways induced by TGF β in a primary airway epithelial cell model of EMT. EMT had dramatic effects on the induction of the innate pathway and the coupling interval of the canonical and noncanonical NF- κ B pathways. Simulation experiments demonstrate that rapid, coordinated cap-independent translation of TRAF-1 and NF- κ B2 is required to reduce the noncanonical pathway coupling interval. Experiments using amantadine confirmed the prediction that TRAF-1 and NF- κ B2/p100 production is mediated by an IRES-dependent mechanism. These data indicate that the epigenetic changes produced by EMT induce dynamic state changes of the innate signaling pathway. Further applications of systems approaches will provide understanding of this complex phenotype through deterministic modeling and multidimensional (genomic and proteomic) profiling.

Show MeSH
Related in: MedlinePlus