Mechanisms of asthma and allergic inflammation
Gene expression profiles in human nasal polyp tissues studied by means of DNA microarray

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Background

Nasal polyposis (NP) is a chronic inflammatory disease of the sinuses. Its pathogenesis is unknown. DNA microarray analysis allows simultaneous measurement of expression of thousands of genes in the same tissue sample and might help to identify gene alterations in various disorders.

Objective

We sought to screen for disease-related genes in NP by using DNA microarrays and to validate the altered expression of selected genes at the mRNA and protein level.

Methods

Expression microarrays containing approximately 10,500 genes were used to compare individual gene profiles of NP samples (n = 10) and normal mucosal samples obtained from sphenoid sinuses in patients undergoing pituitary surgery (n = 4). Four of the 5 most upregulated, and the single most downregulated, genes were retested by means of quantitative RT-PCR and immunohistochemistry in a different set of NP and normal mucosal samples obtained from the ethmoid and sphenoid sinuses.

Results

Compared with normal sinus tissue, 192 genes were upregulated at least 2-fold, and 156 genes were downregulated by at least 50% in NP samples (approximately 3% of genes evaluated). Four of the top 5 overexpressed genes (statherin, 48.0-fold; prolactin-induced protein [PIP], 24.9-fold; lactoferrin, 26.6-fold; and deleted in malignant brain tumor 1 [DMBT1], 30.3-fold) and the most underexpressed gene (Clara cell 10-kd protein [CC10], −20.1-fold) were selected and retested by means of quantitative RT-PCR and immunohistochemical staining. Quantitative RT-PCR and immunohistochemical staining confirmed the differential expression of all except statherin in NP tissue.

Conclusion

DNA microarrays can provide new insight into the possible pathophysiologic processes involved in NP.

Section snippets

Subjects

All studies were conducted by using a human subject research protocol approved by Johns Hopkins Medicine Institutional Review Board.

NP tissues for DNA microarray analysis were obtained from 10 patients with CRS (8 women) between the ages of 27 and 67 years (mean, 51 years). Of these, 3 were identified as allergic on the basis of positive puncture skin test results to a standard panel of aeroallergens. Five of the 10 patients had physician-diagnosed asthma. Two of 10 patients were aspirin

Transcriptional profile in NP

The genes meeting the following criteria were admitted to the list of genes differentially expressed in patients with NP compared with those in healthy patients to ensure a minimal number of false-positive results: (1) the difference of gene expression level between NP and normal tissues was statistically significant, with a P value of less than .05; (2) the expression difference was larger than 2-fold; (3) genes could be called present according to the absolute call criteria in at least half

Discussion

DNA microarrays were used to test more than 10,000 human genes to screen for those in which expression was altered at least 2-fold in NP tissue compared with normal sinus mucosa. This strategy identified 192 upregulated and 156 downregulated genes (Appendixes 4 and 5 in the Journal's Online Repository). The utility of this approach to provide insight into disease pathogenesis is highlighted by the fact that although a few of these genes have been previously implicated in the pathogenesis of NP,

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  • Cited by (0)

    Dr Liu is currently affiliated with the Department of ENT, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Dr Wang is currently affiliated with Merck Research Laboratory, Molecular Profiling, Seattle, Wash.

    Supported by NIH/NIDCR grants DE07652, DE05672, and DE14950 (to F.G.O.).

    Drs Liu and Kim contributed equally to this article.

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