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Published ahead of print on April 5, 2007, doi:10.1165/rcmb.2006-0466OC

Am. J. Respir. Cell Mol. Biol., Volume 37, Number 2, August 2007, 169-185

A more recent version of this article appeared on August 1, 2007
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Submitted on December 19, 2006
Revised on April 4, 2007

Transcriptional Profiling of Mucociliary Differentiation in Human Airway Epithelial Cells

Andrea J Ross1, Lisa A Dailey1, Luisa E Brighton2, and Robert B Devlin1*

1 Clinical Research Branch, Human Studies Division, U.S. Environmental Protection Agency, National Health and Environmental Effects Research Laboratory, Research Triangle Park, NC, USA, 2 Center for Environmental Medicine, Asthma and Lung Biology, University of North Carolina, Chapel Hill, NC, USA

* To whom correspondence should be addressed. E-mail: devlin.robert{at}epa.gov.

When cultured at an air-liquid interface (ALI) in the appropriate medium, primary human airway epithelial cells form a polarized, pseudostratified epithelium composed of ciliated and mucus-secreting cells. This culture system provides a useful tool for the in vitro study of airway epithelial biology and differentiation. We have performed microarray analysis on ALI cultures of human bronchial epithelial cells (HBECs) grown over a 28-day period to identify genes involved in mucociliary differentiation. We identified over 2000 genes that displayed statistically significant two-fold or greater changes in expression during the time course. Of the genes showing the largest increases, many are involved in processes associated with airway epithelial biology, such as cell adhesion, immunity, transport, and cilia formation; however, many novel genes were also identified. We compared our results with data from proteomic analyses of the ciliary axoneme and identified candidate genes that may have roles in cilia formation or function. Gene networks were generated using Ingenuity Pathways Analysis (Ingenuity Systems) to identify signaling pathways involved in mucociliary cell differentiation or function. Networks containing genes involved in TGF{beta}, WNT/{beta}-catenin and epidermal growth factor receptor (EGFR) pathways were identified, suggesting potential roles for these families in airway epithelia. Microarray results were validated by real time RT-PCR for a number of representative genes. This work has provided extensive information about gene expression changes during differentiation of airway epithelial cells, and will be a useful resource for researchers interested in respiratory function, pathology and toxicology.




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