SPDEF is required for mouse pulmonary goblet cell differentiation and regulates a network of genes associated with mucus production
Gang Chen, … , Hans Clevers, Jeffrey A. Whitsett
Gang Chen, … , Hans Clevers, Jeffrey A. Whitsett
Published September 14, 2009
Citation Information: J Clin Invest. 2009;119(10):2914-2924. https://doi.org/10.1172/JCI39731.
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SPDEF is required for mouse pulmonary goblet cell differentiation and regulates a network of genes associated with mucus production

Abstract

Various acute and chronic inflammatory stimuli increase the number and activity of pulmonary mucus-producing goblet cells, and goblet cell hyperplasia and excess mucus production are central to the pathogenesis of chronic pulmonary diseases. However, little is known about the transcriptional programs that regulate goblet cell differentiation. Here, we show that SAM-pointed domain–containing Ets-like factor (SPDEF) controls a transcriptional program critical for pulmonary goblet cell differentiation in mice. Initial cell-lineage–tracing analysis identified nonciliated secretory epithelial cells, known as Clara cells, as the progenitors of goblet cells induced by pulmonary allergen exposure in vivo. Furthermore, in vivo expression of SPDEF in Clara cells caused rapid and reversible goblet cell differentiation in the absence of cell proliferation. This was associated with enhanced expression of genes regulating goblet cell differentiation and protein glycosylation, including forkhead box A3 (Foxa3), anterior gradient 2 (Agr2), and glucosaminyl (N-acetyl) transferase 3, mucin type (Gcnt3). Consistent with these findings, levels of SPDEF and FOXA3 were increased in mouse goblet cells after sensitization with pulmonary allergen, and the proteins were colocalized in goblet cells lining the airways of patients with chronic lung diseases. Deletion of the mouse Spdef gene resulted in the absence of goblet cells in tracheal/laryngeal submucosal glands and in the conducting airway epithelium after pulmonary allergen exposure in vivo. These data show that SPDEF plays a critical role in regulating a transcriptional network mediating the goblet cell differentiation and mucus hyperproduction associated with chronic pulmonary disorders.

Authors

Gang Chen, Thomas R. Korfhagen, Yan Xu, Joseph Kitzmiller, Susan E. Wert, Yutaka Maeda, Alexander Gregorieff, Hans Clevers, Jeffrey A. Whitsett

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Figure 4

SPDEF and FOXA3 are coexpressed and induce Agr2 expression.

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SPDEF and FOXA3 are coexpressed and induce Agr2 expression. Scgb1a1-...
Scgb1a1-rtTA/TRE2-Spdef transgenic mice were treated with doxycycline for 3 days to induce SPDEF in bronchiolar epithelial cells (A and B). SPDEF was colocalized with FOXA3 in nuclei (A) and AGR2 in the cytoplasm (B) of goblet cells as assayed by immunofluorescence microscopy (yellow indicates colocalization of nuclear SPDEF and FOXA3). Luciferase reporter constructs containing the promoter region from the mouse Agr2 (1.6 kb) gene was transfected into primary sheep tracheal epithelial cells. Synergistic activation of Agr2 promoter was observed when SPDEF and FOXA3 were cotransfected (C). Endogenous Agr2 mRNA expression was induced by transfection with both SPDEF and FOXA3 expression plasmids in the mouse lung epithelial cell line MLE15 (D). Results were expressed as mean ± SD of 3 independent experiments. *P < 0.02; **P < 0.01 versus control constructs. Scale bars: 25 μm.