Epidermal Growth Factor Removal or Tyrphostin AG1478 Treatment Reduces Goblet Cells & Mucus Secretion of Epithelial Cells from Asthmatic Children Using the Air-Liquid Interface Model

PLoS One. 2015 Jun 9;10(6):e0129546. doi: 10.1371/journal.pone.0129546. eCollection 2015.

Abstract

Rationale: Epithelial remodelling in asthma is characterised by goblet cell hyperplasia and mucus hypersecretion for which no therapies exist. Differentiated bronchial air-liquid interface cultures from asthmatic children display high goblet cell numbers. Epidermal growth factor and its receptor have been implicated in goblet cell hyperplasia.

Objectives: We hypothesised that EGF removal or tyrphostin AG1478 treatment of differentiating air-liquid interface cultures from asthmatic children would result in a reduction of epithelial goblet cells and mucus secretion.

Methods: In Aim 1 primary bronchial epithelial cells from non-asthmatic (n = 5) and asthmatic (n = 5) children were differentiated under EGF-positive (10 ng/ml EGF) and EGF-negative culture conditions for 28 days. In Aim 2, cultures from a further group of asthmatic children (n = 5) were grown under tyrphostin AG1478, a tyrosine kinase inhibitor, conditions. All cultures were analysed for epithelial resistance, markers of differentiation using immunocytochemistry, ELISA for MUC5AC mucin secretion and qPCR for MUC5AC mRNA.

Results: In cultures from asthmatic children the goblet cell number was reduced in the EGF negative group (p = 0.01). Tyrphostin AG1478 treatment of cultures from asthmatic children had significant reductions in goblet cells at 0.2 μg/ml (p = 0.03) and 2 μg/ml (p = 0.003) as well as mucus secretion at 2 μg/ml (p = 0.04).

Conclusions: We have shown in this preliminary study that through EGF removal and tyrphostin AG1478 treatment the goblet cell number and mucus hypersecretion in differentiating air-liquid interface cultures from asthmatic children is significantly reduced. This further highlights the epidermal growth factor receptor as a potential therapeutic target to inhibit goblet cell hyperplasia and mucus hypersecretion in asthma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Air*
  • Asthma / genetics
  • Asthma / pathology*
  • Bronchi / pathology
  • Cell Count
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Child
  • Child, Preschool
  • Electric Impedance
  • Epidermal Growth Factor / isolation & purification*
  • Female
  • Goblet Cells / drug effects
  • Goblet Cells / metabolism*
  • Humans
  • Infant
  • Male
  • Models, Biological*
  • Mucin 5AC / genetics
  • Mucin 5AC / metabolism*
  • Quinazolines / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Tyrphostins / pharmacology*

Substances

  • Mucin 5AC
  • Quinazolines
  • RNA, Messenger
  • Tyrphostins
  • RTKI cpd
  • Epidermal Growth Factor

Grants and funding

Northern Ireland Chest Heart and Stroke Association (grant number 200942) provided funding for this study. The funders had no involvement in the design, data collection, analysis, decision to publish or preparation of the manuscript.