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 1

SPDEF caused differentiation of goblet cells from Clara cells.

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SPDEF caused differentiation of goblet cells from Clara cells. (A) Adult...
(A) Adult Scgb1a1-rtTA/Otet-Cre mice were mated to R26R mice. (B) To permanently label Clara cells, doxycycline (Dox) was administered from days 12 to 17 during i.p. sensitization with OVA. Mice were sacrificed either before receiving the first i.n. sensitization (day 24) to assess Clara cell labeling with β-gal or after the second i.n. sensitization (day 29) with OVA to induce goblet cell hyperplasia. (C) Before pulmonary OVA sensitization, β-gal was expressed in nonciliated epithelial cells, as shown by its exclusion from FOXJ1-expressing cells and colocalization with CCSP. After sensitization, β-gal was detected (white pseudocolor) in most goblet cells, identified by MUC5AC, indicating their derivation from Clara cells. MUC5AC, β-gal, and FOXJ1 were not colocalized. (D) Scgb1a1-rtTA/TRE2-Spdef mice were treated 3 days with or without doxycycline. SPDEF induced goblet cell differentiation as detected by Alcian blue (AB) and by changes in cell morphology. SPDEF staining decreased 4 and 8 days after withdrawal of doxycycline, at which time goblet cell differentiation was substantially resolved. CCSP was decreased in the conducting airway epithelium 3 days after induction of SPDEF. 4 to 8 days after withdrawal of doxycycline, CCSP staining was restored. Inserts show higher magnifications of the regions indicated by arrows. Scale bars: 25 μm.