SPDEF regulates goblet cell hyperplasia in the airway epithelium
Kwon-Sik Park, … , Gang Chen, Jeffrey A. Whitsett
Kwon-Sik Park, … , Gang Chen, Jeffrey A. Whitsett
Published April 2, 2007
Citation Information: J Clin Invest. 2007;117(4):978-988. https://doi.org/10.1172/JCI29176.
View: Text | PDF
Research Article Pulmonology

SPDEF regulates goblet cell hyperplasia in the airway epithelium

Abstract

Goblet cell hyperplasia and mucous hypersecretion contribute to the pathogenesis of chronic pulmonary diseases including cystic fibrosis, asthma, and chronic obstructive pulmonary disease. In the present work, mouse SAM pointed domain-containing ETS transcription factor (SPDEF) mRNA and protein were detected in subsets of epithelial cells lining the trachea, bronchi, and tracheal glands. SPDEF interacted with the C-terminal domain of thyroid transcription factor 1, activating transcription of genes expressed selectively in airway epithelial cells, including Sftpa, Scgb1a1, Foxj1, and Sox17. Expression of Spdef in the respiratory epithelium of adult transgenic mice caused goblet cell hyperplasia, inducing both acidic and neutral mucins in vivo, and stainined for both acidic and neutral mucins in vivo. SPDEF expression was increased at sites of goblet cell hyperplasia caused by IL-13 and dust mite allergen in a process that was dependent upon STAT-6. SPDEF was induced following intratracheal allergen exposure and after Th2 cytokine stimulation and was sufficient to cause goblet cell differentiation of Clara cells in vivo.

Authors

Kwon-Sik Park, Thomas R. Korfhagen, Michael D. Bruno, Joseph A. Kitzmiller, Huajing Wan, Susan E. Wert, Gurjit K. Khurana Hershey, Gang Chen, Jeffrey A. Whitsett

×

Figure 4

SPDEF interacts with TTF-1 via the C-terminal domain of TTF-1.

Options: View larger image (or click on image) Download as PowerPoint
SPDEF interacts with TTF-1 via the C-terminal domain of TTF-1. (A) Mamma...
(A) Mammalian 2-hybrid assay was performed using the luciferase reporter pG5-luc and pACT and pBIND as described in Methods. Full-length TTF-1 and a series of TTF-1 deletion mutants (see Supplemental Table 1) were inserted to pBIND vector. Recombinant plasmids were cotransfected with the pG5-luc plasmids, and their activity was compared with that of cells transfected with pACT-SPDEF plus pBIND–TTF-1. Values are mean ± SD (n = 3). Assays were repeated 3 times with similar results. HD, homeodomain. (B) GST pulldown assays were performed with GST-SPDEF that was immobilized on glutathione-sepharose beads. Protein extracts were prepared from HeLa cells transiently transfected with the expression plasmids encoding for 3XFLAG–TTF-1, 3XFLAGΔ14, and 3XFLAGΔ3 as described in Methods. The extracts were incubated with GST or GST-SPDEF. Both GST and GST-SPDEF beads were washed several times before boiling, run on 10% SDS-polyacrylamide gels, and analyzed by immunoblot using a monoclonal antibody that recognizes the FLAG sequences.