Blocking notch3 signaling abolishes MUC5AC production in airway epithelial cells from individuals with asthma

Andrew T. Reid, Kristy S. Nichol, Punnam Chander Veerati, Fatemeh Moheimani, Anthony Kicic, Stephen M. Stick, Nathan W. Bartlett, Chris L. Grainge, Peter A.B. Wark, Philip M. Hansbro, Darryl A. Knight

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3 Citations (Scopus)

Abstract

In asthma, goblet cell numbers are increased within the airway epithelium, perpetuating the production of mucus that is more difficult to clear and results in airway mucus plugging. Notch1, Notch2, or Notch3, or a combination of these has been shown to influence the differentiation of airway epithelial cells. How the expression of specific Notch isoforms differs in fully differentiated adult asthmatic epithelium and whether Notch influences mucin production after differentiation is currently unknown. We aimed to quantify different Notch isoforms in the airway epithelium of individuals with severe asthma and to examine the impact of Notch signaling on mucin MUC5AC. Human lung sections and primary bronchial epithelial cells from individuals with and without asthma were used in this study. Primary bronchial epithelial cells were differentiated at the air-liquid interface for 28 days. Notch isoform expression was analyzed by Taqman quantitative PCR. Immunohistochemistry was used to localize and quantify Notch isoforms in human airway sections. Notch signaling was inhibited in vitro using dibenzazepine or Notch3-specific siRNA, followed by analysis of MUC5AC. NOTCH3 was highly expressed in asthmatic airway epithelium compared with nonasthmatic epithelium. Dibenzazepine significantly reduced MUC5AC production in air-liquid interface cultures of primary bronchial epithelial cells concomitantly with suppression of NOTCH3 intracellular domain protein. Specific knockdown using NOTCH3 siRNA recapitulated the dibenzazepine-induced reduction in MUC5AC. We demonstrate that NOTCH3 is a regulator of MUC5AC production. Increased NOTCH3 signaling in the asthmatic airway epithelium may therefore be an underlying driver of excess MUC5AC production.

Original languageEnglish
Pages (from-to)513-523
Number of pages11
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume62
Issue number4
DOIs
Publication statusPublished - 1 Jan 2020

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    Reid, A. T., Nichol, K. S., Veerati, P. C., Moheimani, F., Kicic, A., Stick, S. M., Bartlett, N. W., Grainge, C. L., Wark, P. A. B., Hansbro, P. M., & Knight, D. A. (2020). Blocking notch3 signaling abolishes MUC5AC production in airway epithelial cells from individuals with asthma. American Journal of Respiratory Cell and Molecular Biology, 62(4), 513-523. https://doi.org/10.1165/rcmb.2019-0069OC