Persistent induction of goblet cell differentiation in the airways: Therapeutic approaches

Reid, Andrew T; Veerati, Punnam Chander; Gosens, Reinoud; Bartlett, Nathan W.; Wark, Peter; Grainge, Christopher; Stick, Stephen M.; Kicic, Anthony; Moheimani, Fatemeh; Hansbro, Philip M.; Knight, Darryl A.

doi:10.1016/j.pharmthera.2017.12.009

Cited by 27 publications

(25 citation statements)

References 239 publications

(217 reference statements)

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“…Thus, while glucocorticoids can modulate such processes, details of activation/repression combined with timing and location may be critical to appreciate physiological roles. This may integrate with increased expression of genes involved in cytokine signaling, PI3K, Akt, FOXO, mTOR pathways and others (this study, [24]), and may collectively impact on wound repair, healing and/or alternatively on unwanted remodeling [69][70][71].…”

Section: Discussionmentioning

confidence: 94%

Glucocorticoid-Driven Transcriptomes in Human Airway Epithelial Cells: Commonalities, Differences and Functional Insight from Cell Lines and Primary Cells

Mostafa

Rider

Newton

2019

A61. Epithelial Biology

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RATIONALE Glucocorticoids are stress response hormones that act on the glucocorticoid receptor (GR) to execute effects, including repression of inflammatory gene expression. The molecular mechanisms for this process are still elusive, indeed controversial. This is, to a large extent, due the knowledge gap in understanding the functional impacts of the key glucocorticoid‐modulated genes in relevant tissues. Identification of such genes may be problematic because the effects of glucocorticoids on gene expression are context‐dependent and show variable effects between different cell types. This confounds the identification of key gene expression features within the glucocorticoid‐response. The current study addresses this by comparing the glucocorticoid‐driven transcriptomes in 3 variants of human airway epithelial cells, a critical player in airway inflammation and physiological responses to glucocorticoid. These include two airway epithelial cell lines, pulmonary type II A549 and bronchial epithelial BEAS‐2B cells, and primary human bronchial epithelial (HBE) cells. METHODS Gene expression profiling of RNA extracted from A549, BEAS‐2B and HBE cells following budesonide treatment for 6 h was performed using Affymetrix PrimeView microarrays. Genes showing significant induction (fold ≥ 2, P ≤ 0.05) or repression (fold ≤ 0.5, P ≤ 0.05) compared to untreated cells, in any of the cell types were used for further analyses. Representative genes were validated using qPCR. Gene ontology and Ingenuity Pathway Analysis (IPA) were performed using differentially regulated genes in each cell variant. RESULTS Using stringent (fold ≥ 2 or ≤ 0.5, P ≤ 0.05) cutoff criteria, only 17 and 8 of genes induced or repressed, respectively, by glucocorticoid were common to all 3 epithelial cell variants. While a major fraction of the significantly modulated genes was apparently unique to each cell variant, hierarchical clustering revealed greater commonality than was suggested by the stringent cut‐off. Applying less stringent cut‐offs (fold ≥ 1.25 or ≤ 0.8) within the pool of genes identified by stringent cut‐off revealed 93 induced and 82 repressed genes that were in common among all 3 cell variants. Validation of 52 budesonide‐induced genes using qPCR confirmed, for most of the genes, the grouping scheme determined by applying the less stringent cut‐off. Gene ontology (GO) analysis of the genes induced in common showed enrichment of transcriptional control, proliferation/apoptosis and signaling terms. Equally, the GO term growth factor activity was profoundly enriched with budesonide‐repressed genes across all variants. CONCLUSIONS The current comparative transcriptome analysis revealed a surprisingly large variance among 3 variants of airway epithelial cells in response to glucocorticoids. Despite this, the common gene expression features obtained by such analysis represent unbiased listing of potentially key molecular players in glucocorticoid response. While many of these genes, and the associated pathways, are consistent with anti‐infl...

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Section: Discussionmentioning

confidence: 94%

Glucocorticoid-Driven Transcriptomes in Human Airway Epithelial Cells: Commonalities, Differences and Functional Insight from Cell Lines and Primary Cells

Mostafa

Rider

Newton

2019

A61. Epithelial Biology

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“…The use of VPA to activate the Notch pathway is widely stablished in different cellular types, such as Clara cell and carcinoma models. Although the effect of this molecule is highly cell specific, a clear role of VPA in activating the Notch pathway and blocking Clara cell transdifferentiation has been reported [10]. In addition, we also determined the transnucleation of the Notch intracellular domain both in Pneumocystis infected and noninfected animals.…”

Section: Discussionmentioning

confidence: 98%

“…Differentiation from Clara to goblet cells in postnatal development is negatively regulated by the Notch signaling pathway [9]. The role of Notch pathway in regulating transformation of Clara to mucous cells is well established[10], and can be reversed by using Notch antagonists that induce an increase in goblet cells in human epithelial cells[11]. Notch is a master regulatory circuit involved in cellular proliferation, differentiation and apoptosis[12].…”

Section: Introductionmentioning

confidence: 99%

Primary infection by Pneumocystis induces Notch-independent Clara cell mucin production in rat distal airways

et al. 2019

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Clara cells are the main airway secretory cells able to regenerate epithelium in the distal airways through transdifferentiating into goblet cells, a process under negative regulation of the Notch pathway. Pneumocystis is a highly prevalent fungus in humans occurring between 2 and 5 months of age, a period when airways are still developing and respiratory morbidity typically increases. Pneumocystis induces mucus hyperproduction in immunocompetent host airways and whether it can stimulate Clara cells is unknown. Markers of Clara cell secretion and Notch1 activation were investigated in lungs of immunocompetent rats at 40, 60, and 80 days of age during Pneumocystis primary infection with and without Valproic acid (VPA), a Notch inducer. The proportion of rats expressing mucin increased in Pneumocystis -infected rats respect to controls at 60 and 80 days of age. Frequency of distal airways Clara cells was maintained while mRNA levels for the mucin-encoding genes Muc5B and Muc5ac in lung homogenates increased 1.9 and 3.9 times at 60 days of infection ( P . = 0.1609 and P . = 0.0001, respectively) and protein levels of the Clara cell marker CC10 decreased in the Pneumocystis -infected rats at 60 and 80 days of age ( P . = 0.0118 & P. = 0.0388). CC10 and Muc5b co-localized in distal airway epithelium of Pneumocystis -infected rats at day 60. Co-localization of Muc5b and Ki67 as marker of mitosis in distal airways was not observed suggesting that Muc5b production by Clara cells was independent of mitosis. Notch levels remained similar and no transnucleation of activated Notch associated to Pneumocystis infection was detected. Unexpectedly, mucus was greatly increased at day 80 in Pneumocystis -infected rats receiving VPA suggesting that a Notch-independent mechanism was triggered. Overall, data suggests a Clara to goblet cell transdifferentiation mechanism induced by Pneumocystis and independent of Notch.

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“…Mucus overproduction is one of many irregularities typically associated with inflammatory airway diseases asthma, chronic obstructive pulmonary disease (COPD) and bronchiectasis. When coupled with increases in viscosity, it is linked to defective mucociliary clearance and the formation of mucus plugs that cause airflow obstruction (1,2). Two major gel-forming mucins exist in the airway mucus layer, MUC5AC and MUC5B.…”

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confidence: 99%

Blocking Notch3 Signaling Abolishes MUC5AC Production in Airway Epithelial Cells from Individuals with Asthma

Reid

Nichol

Veerati

et al. 2020

Am J Respir Cell Mol Biol

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Author Contributions: A.R. designed and performed experiments, analyzed results, and wrote the manuscript. K.N. and P.V. helped with methodology, performed experiments and analyzed results. F.M., A.K., S.S. and N.B. helped design experiments and reviewed the manuscript. P.W. provided primary human bronchial epithelial cells and reviewed the manuscript. C.G. and P.H. helped conceptualize design of the project, helped interpret data and reviewed the manuscript. D.K. helped conceptualize project design, supervised the project, interpreted results and reviewed the manuscript.

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Persistent induction of goblet cell differentiation in the airways: Therapeutic approaches

Cited by 27 publications

References 239 publications

Glucocorticoid-Driven Transcriptomes in Human Airway Epithelial Cells: Commonalities, Differences and Functional Insight from Cell Lines and Primary Cells

Glucocorticoid-Driven Transcriptomes in Human Airway Epithelial Cells: Commonalities, Differences and Functional Insight from Cell Lines and Primary Cells

Primary infection by Pneumocystis induces Notch-independent Clara cell mucin production in rat distal airways

Blocking Notch3 Signaling Abolishes MUC5AC Production in Airway Epithelial Cells from Individuals with Asthma

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