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

Abstract

RationaleEpithelial 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.ObjectivesWe 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.MethodsIn Aim 1 primary bronchial epithelial cells from non-asthmatic (n = 5) and asthmatic (n = 5) children were differentiated under EGF-positive (10ng/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.ResultsIn 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 0.2μg/ml (p = 0.03) and 2μg/ml (p = 0.003) as well as mucus secretion at 2μg/ml (p = 0.04).ConclusionsWe 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.

Nononcogenic restoration of the intestinal barrier by E. coli–delivered human EGF

“…Moreover, EGF-EcN was shown to attenuate goblet cell depletion and mucin suppression in the insulted gut environment in the present study. Signaling activation by EGF has been reported to stimulate goblet cell growth and secretion in the mucosal epithelia (51,52). In addition to the protective actions of the recombinant probiotic bacteria in the colon, EGF-EcN alleviated ulcerative injuries in the small intestine in similar ways.…”

Impact of atopy, asthma, and asthma treatment on nasal epithelial wound healing

“…To determine the role of the EGF pathway on wound healing, EGF (25ng/ml, StemCell) 13 was added to the basolateral media 7 days prior to wounding and replenished every two days with media changes. In separate experiments EGF was neutralized by adding the specific EGF receptor blocker, Erlotinib HCl (2μg/ml) (OSI-744, Shelleckchem) 14 , to the basolateral media 7 days prior to wounding. Media with inhibitor was replenished every two days.…”

Characterizing well-differentiated culture of primary human nasal epithelial cells for use in wound healing assays

“…Erlotinib HCl (2 μg/mL)(OSI-744, Shelleckchem) [27] was added to the basolateral media 7 days prior to wounding and replenished every two days when media was changed.…”