Airway Hyperresponsiveness in Asthma Model Occurs Independently of Secretion of β1 Integrins in Airway Wall and Focal Adhesions Proteins Down Regulation

Álvarez-Santos, Mayra D.; Carbajal, Verónica; Tellez-Jiménez, Olivia; Martínez-Cordero, E; Ruiz, Vı́ctor; Hernández-Pando, Rogelio; Lascuraín, Ricardo; Santibañez-Salgado, Alfredo; Bazán‐Perkins, Blanca

doi:10.1002/jcb.25536

Cited by 3 publications

(3 citation statements)

References 49 publications

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“…ITGB1 encodes an integrin beta subunit that functions as receptor for fibronectin; along with the increased expression of fibronectin, this would result in enhanced functional consequences for the airway. Interestingly, a recent study in a guinea pig model found that ITGB1 expression was increased in asthma 57 . Vinculin ( VCL ), that has a role in regulating cell–matrix adhesion, including the assembly, turnover, and strength of focal adhesions, as well as the transmission of force by these cellular structures, was downregulated in myofibroblasts.…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic changes during TGF-β-mediated differentiation of airway fibroblasts to myofibroblasts

Walker

Heydet

Veldre

et al. 2019

Sci Rep

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Asthma is the most common chronic lung disease in children and young adults worldwide. Airway remodelling (including increased fibroblasts and myofibroblasts in airway walls due to chronic inflammation) differentiates asthmatic from non-asthmatic airways. The increase in airway fibroblasts and myofibroblasts occurs via epithelial to mesenchymal transition (EMT) where epithelial cells lose their tight junctions and are transdifferentiated to mesenchymal cells, with further increases in myofibroblasts occurring via fibroblast-myofibroblast transition (FMT). Transforming growth factor (TGF)-β is the central EMT- and FMT-inducing cytokine. In this study, we have used next generation sequencing to delineate the changes in the transcriptome induced by TGF-β treatment of WI-38 airway fibroblasts in both the short term and after differentiation into myofibroblasts, to gain an understanding of the contribution of TGF-β induced transdifferentiation to the asthmatic phenotype. The data obtained from RNAseq analysis was confirmed by quantitative PCR (qPCR) and protein expression investigated by western blotting. As expected, we found that genes coding for intermediates in the TGF-β signalling pathways (SMADs) were differentially expressed after TGF-β treatment, SMAD2 being upregulated and SMAD3 being downregulated as expected. Further, genes involved in cytoskeletal pathways (FN1, LAMA, ITGB1) were upregulated in myofibroblasts compared to fibroblasts. Importantly, genes that were previously shown to be changed in asthmatic lungs (ADAMTS1, DSP, TIMPs, MMPs) were similarly differentially expressed in myofibroblasts, strongly suggesting that TGF-β mediated differentiation of fibroblasts to myofibroblasts may underlie important changes in the asthmatic airway. We also identified new intermediates of signalling pathways (PKB, PTEN) that are changed in myofibroblasts compared to fibroblasts. We have found a significant number of genes that are altered after TGF-β induced transdifferentiation of WI-38 fibroblasts into myofibroblasts, many of which were expected or predicted. We also identified novel genes and pathways that were affected after TGF-β treatment, suggesting additional pathways are activated during the transition between fibroblasts and myofibroblasts and may contribute to the asthma phenotype.

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Section: Discussionmentioning

confidence: 99%

Transcriptomic changes during TGF-β-mediated differentiation of airway fibroblasts to myofibroblasts

Walker

Heydet

Veldre

et al. 2019

Sci Rep

View full text Add to dashboard Cite

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“…The myofilament-localized ILK isoform contains a series of ankyrin regions similar to those in the PP1c subunit of MLCP, and this isoform may interact with MLC20 (Lynch et al, 1999). However, its role in hypercontractility and hyperresponsiveness is unclear, because ILK expression has been demonstrated to be reduced in experimental asthma (Álvarez-Santos et al, 2016).…”

Section: Integrin-linked Kinase (Ilk) As Regulator Of Mlcp In Airway mentioning

confidence: 99%

“…In cultured A7r5 rat embryonic aorta smooth muscle cells, ROCK-mediated phosphorylation of MYPT1 at Thr 695 is similar to that of phosphorylation of human MYPT1 at Thr 696 , and phosphorylation of rat MYPT1 at Thr 850 is similar to that of phosphorylation of human MYPT1 at Thr 853 ( Murányi et al, 2005 ). In a guinea pig model of asthma and in airway smooth muscle from asthma patients, increased phosphorylation of MYPT1 occurs at Thr 696 ( Álvarez-Santos et al, 2016 ; Yoo et al, 2017 ). In addition, phosphorylation of MYPT1 at Thr 696 by ROCK in bovine airway smooth muscle inhibits MLCP activity, although Thr 853 phosphorylation does not alter its phosphatase activity ( Gao et al, 2017 ).…”

Section: Rho-associated Kinase (Rock) Is a Key Regulator Of Airway Smmentioning

confidence: 99%

Regulation of Myosin Light-Chain Phosphatase Activity to Generate Airway Smooth Muscle Hypercontractility

et al. 2020

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Smooth muscle is a central structure involved in the regulation of airway tone. In addition, it plays an important role in the development of some pathologies generated by alterations in contraction, such as hypercontractility and the airway hyperresponsiveness observed in asthma. The molecular processes associated with smooth muscle contraction are centered around myosin light chain (MLC) phosphorylation, which is controlled by a balance in the activity of myosin light-chain kinase (MLCK) and myosin light-chain phosphatase (MLCP). MLCK activation depends on increasing concentrations of intracellular Ca 2+ , while MLCP activation is independent of Ca 2+. MLCP contains a phosphatase subunit (PP1c) that is regulated through myosin phosphatase target subunit 1 (MYPT1) and other subunits, such as glycogenassociated regulatory subunit and myosin-binding subunit 85 kDa. Interestingly, MLCP inhibition may contribute to exacerbation of smooth muscle contraction by increasing MLC phosphorylation to induce hypercontractility. Many pathways inhibiting MLCP activity in airway smooth muscle have been proposed and are focused on inhibition of PP1c, inhibitory phosphorylation of MYPT1 and dissociation of the PP1c-MYPT1 complex.

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The Utility of Resolving Asthma Molecular Signatures Using Tissue-Specific Transcriptome Data

Ghosh

Ding

Bernstein

et al. 2020

G3 Genes|Genomes|Genetics

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An integrative analysis focused on multi-tissue transcriptomics has not been done for asthma. Tissue-specific DEGs remain undetected in many multi-tissue analyses, which influences identification of disease-relevant pathways and potential drug candidates. Transcriptome data from 609 cases and 196 controls, generated using airway epithelium, bronchial, nasal, airway macrophages, distal lung fibroblasts, proximal lung fibroblasts, CD4+ lymphocytes, CD8+ lymphocytes from whole blood and induced sputum samples, were retrieved from Gene Expression Omnibus (GEO). Differentially regulated asthma-relevant genes identified from each sample type were used to identify (a) tissue-specific and tissue-shared asthma pathways, (b) their connection to GWAS-identified disease genes to identify candidate tissue for functional studies, (c) to select surrogate sample for invasive tissues, and finally (d) to identify potential drug candidates via connectivity map analysis. We found that inter-tissue similarity in gene expression was more pronounced at pathway/functional level than at gene level with highest similarity between bronchial epithelial cells and lung fibroblasts, and lowest between airway epithelium and whole blood samples. Although public-domain gene expression data is limited by inadequately annotated per-sample demographic and clinical information which limited the analysis, our tissue-resolved analysis clearly demonstrated relative importance of unique and shared asthma pathways, At the pathway level, IL-1b signaling and ERK signaling were significant in many tissue types, while Insulin-like growth factor and TGF-beta signaling were relevant in only airway epithelial tissue. IL-12 (in macrophages) and Immunoglobulin signaling (in lymphocytes) and chemokines (in nasal epithelium) were the highest expressed pathways. Overall, the IL-1 signaling genes (inflammatory) were relevant in the airway compartment, while pro-Th2 genes including IL-13 and STAT6 were more relevant in fibroblasts, lymphocytes, macrophages and bronchial biopsies. These genes were also associated with asthma in the GWAS catalog. Support Vector Machine showed that DEGs based on macrophages and epithelial cells have the highest and lowest discriminatory accuracy, respectively. Drug (entinostat, BMS-345541) and genetic perturbagens (KLF6, BCL10, INFB1 and BAMBI) negatively connected to disease at multi-tissue level could potentially repurposed for treating asthma. Collectively, our study indicates that the DEGs, perturbagens and disease are connected differentially depending on tissue/cell types. While most of the existing literature describes asthma transcriptome data from individual sample types, the present work demonstrates the utility of multi-tissue transcriptome data. Future studies should focus on collecting transcriptomic data from multiple tissues, age and race groups, genetic background, disease subtypes and on the availability of better-annotated data in the public domain.

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Airway Hyperresponsiveness in Asthma Model Occurs Independently of Secretion of β1 Integrins in Airway Wall and Focal Adhesions Proteins Down Regulation

Cited by 3 publications

References 49 publications

Transcriptomic changes during TGF-β-mediated differentiation of airway fibroblasts to myofibroblasts

Transcriptomic changes during TGF-β-mediated differentiation of airway fibroblasts to myofibroblasts

Regulation of Myosin Light-Chain Phosphatase Activity to Generate Airway Smooth Muscle Hypercontractility

The Utility of Resolving Asthma Molecular Signatures Using Tissue-Specific Transcriptome Data

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