Triological Society Candidate Thesis
Differential Gene Expression in Cholesteatoma by DNA Chip Analysis
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).
The authors have no other funding, financial relationships, or conflicts of interest to disclose.
Send correspondence to John D. Macias, MD, Macias Otology, P.C., 1515 North 9th Street, Suite B, Phoenix, AZ 85006. E-mail: firstname.lastname@example.org
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