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College, Institute, or Department
MD-PhD Scholars Program
John D. Dickinson
Elizabeth B. Staab
Inflammatory airway diseases (e.g. COPD and asthma) are associated with mucous cell metaplasia and mucin hypersecretion, resulting in symptoms such as shortness of breath and cough. However, how secretory cells remove excess mucin granules is poorly understood. Previous research suggests that intracellular degradation pathways, such as autophagy, are involved in the degradation of mucin granules during resolution of mucous cell metaplasia. We thus hypothesized that the elimination of excess mucin granules is dependent on lysosome-mediated degradation in airway secretory cells. Calu-3 cells, an airway epithelial cell line containing abundant mucin granules, were treated with inhibitors of lysosome acidification (Bafilomycin A1) and lysosome enzyme activity (Pepstatin E64d). We found statistically significant increases in the levels of secretory mucin, MUC5AC, by mucin blot, suggesting that the lysosome mediates the elimination of mucin granules. In addition, by immunoblot we observed an increase in the autophagosome markers, LC3-II and SQSTM1, with lysosome inhibition using Bafilomycin A1, indicating an accumulation of autophagosomes and a role for autophagy in the degradation of mucin granules. However, after transfecting Calu-3 cells with a ubiquitin-hemagglutinin tag plasmid to examine the role of the proteasome in the degradation of mucin granules, we observed that our transfection efficiency was low, making it difficult to detect the hemagglutinin epitope by immunoblots. Nevertheless, we found that MUC5AC levels preliminarily increase with the inhibition of the proteasome using MG-132, suggesting a potential role for the proteasome in the degradation of mucin granules. Thus, we can conclude that inhibition of the lysosome increases MUC5AC levels, demonstrating that the lysosome mediates the degradation of mucin granules in airway secretory cells. In addition, while we were not able to conclude that the proteasome is involved in the degradation of mucin granules with certainty, our preliminary data suggests that it is possible that the ubiquitin-proteasome system is involved in the degradation of mucin due to the observed increase in MUC5AC levels with MG-132.
MUC5AC, Mucin Degradation, Lysosome, Proteasome
Sillaste, Erik A.; Thomes, Paul G.; Staab, Elizabeth B.; and Dickinson, John D., "Degradation of Airway Secretory Cell Mucin Granules Is Dependent on Lysosome Activity" (2022). Posters: 2022 Summer Undergraduate Research Program. 38.