TIMECOURSE OF TH2 CYTOKINE-INDUCED GOBLET CELL HYPERPLASIA IN THE EPIAIRWAY™ IN VITRO HUMAN AIRWAY MODEL.

INTRODUCTION: Goblet cell hyperplasia and associated airway re-modeling contribute significantly to the pathogenesis of airway diseases such as asthma, rhinitis and COPD. TH2 cytokines including IL-4 and IL-13 are known to induce goblet cell hyperplasia in various in vivo animal models and in vitro airway cell model systems. However, the timecourse of induction and recovery of this effect are not well characterized. In the current work, well-differentiated organotypic in vitro human airway models (EpiAirway and EpiAirway-FT) were utilized to further characterize induction of goblet cell hyperplasia and gene expression changes caused by TH2 cytokines. METHODS: Fully differentiated organotypic models (EpiAirway, EpiAirway-FT, Figures 1-3) were exposed to IL-4 or IL-13 at various doses over the course of a 6 day period. Goblet cell hyperplasia was evaluated in H&E stained paraffin sections and by PAS staining. Changes in mucin signaling pathway gene expression were evaluated by qPCR arrays (1-10). RESULTS: IL-4 and IL-13 were each capable of inducing goblet cell hyperplasia within 4 to 6 days at 1 ng/ml. IL-13 was the more potent inducer of this effect (Figures 4-5). Numerous changes in mucin and associated signaling pathway gene expression, including induction of ALOX15, CLCA1, MUC5AC, SPDEF and TFF3, were observed in response to the TH2 cytokine treatment (Tables 1-2). Goblet cell hyperplasia was only slight reversed at 6 days following removal of TH2 cytokine (Figure 6). CONCLUSIONS: Organotypic in vitro human airway models appear to be useful for long-term goblet cell hyperplasia induction and remodeling experiments. This system will provide further understanding of the mechanisms of goblet cell hyperplasia and airway remodeling at the molecular level, and should also prove useful for development of novel therapeutic approaches to alleviation of airway diseases associated with TH2 imbalances.

ALOX15, Airway diseases, Alcian blue, Alleviation of airway, CLCA1, EpiAirway, EpiAirway-FT, Gene expression, Goblet cell, Goblet cell hyperplasia, MUC5AC, Mucin signaling, PAS staining, SPEDF, TFF3, TH2 cytokine treatment, TH2 imbalances

IL-13, IL-4