Funding was received by Banner Good Samaritan Medical Center, the Nathan Cummings Foundation with the support of Sheila and Michael Zuieback, the Arizona Community Foundation, the National Organization for Hearing Research Foundation, and the EAR Foundation of Arizona. Toray Industries provided material and financial support for this research (3D-Gene Competition Award for Microarray Analysis).
Triological Society Candidate Thesis
Differential Gene Expression in Cholesteatoma by DNA Chip Analysis
Article first published online: 13 MAY 2013
Copyright © 2013 The American Laryngological, Rhinological and Otological Society, Inc.
Volume 123, Issue Supplement S5, pages S1–S21, October 2013
How to Cite
Macias, J. D., Gerkin, R. D., Locke, D. and Macias, M. P. (2013), Differential Gene Expression in Cholesteatoma by DNA Chip Analysis. The Laryngoscope, 123: S1–S21. doi: 10.1002/lary.24176
The authors have no other funding, financial relationships, or conflicts of interest to disclose.
- Issue published online: 23 SEP 2013
- Article first published online: 13 MAY 2013
- Manuscript Accepted: 3 APR 2013
- Manuscript Revised: 20 MAR 2013
- Manuscript Received: 21 FEB 2013
- gene expression;
In contrast to normal epithelium, the desquamating stratified squamous epithelium of temporal bone cholesteatoma characteristically exhibits sustained hyperproliferative growth and a capacity for bone erosion. We conducted genome-wide microarray analyses to determine the molecular nature of cholesteatoma's biological processes and identify disease-associated, altered gene activity. We tested the hypothesis that genes contributing to the pathophysiology of cholesteatoma are differentially expressed compared to control tissue.
Prospective experimental analysis.
Using new, enhanced microarray platforms and well-annotated human transcriptome probes, we measured global gene expression levels in surgical specimens of cholesteatoma and in the corresponding normal postauricular skin in four patients. Genes of interest were verified by quantitative real time reverse transcriptase polymerase chain reaction analyses using cholesteatoma and postauricular sample pairs (n = 13). External auditory canal skin from six additional patients was also evaluated as a normal control. Immunohistochemistry detected protein expression in tissue sections and the cells involved.
DNA chip analyses identified 282 differentially expressed genes in cholesteatoma compared to control samples. Of these, 104 genes were upregulated and 178 were downregulated. Ontological classifications indicate relationships to cellular processes including receptor binding, cell communication and motion, vitamin metabolism, and cytokine-mediated inflammation. Based on potential involvement in disease pathology, 10 genes were selected and independently verified by quantitative polymerase chain reaction. Immunohistochemical detection of transcobalamin-1 and CCL27 implicates cholesteatoma keratinocytes and dermal endothelial cells as contributors in disease processes.
We present a comprehensive, human genome-wide survey of disease-associated gene expression that extends the public database and provides new evidence for molecular mechanisms involved in cholesteatoma pathology. Laryngoscope, 123:S1–S21, 2013