Alterations of epithelial stem cell marker patterns in human diabetic corneas and effects of c-met gene therapy
| Autor: | Saghizadeh, M., Soleymani, S., Harounian, A., Bhakta, B., Troyanovsky, S. M., Brunken, W. J., Pellegrini, G., Alexander Ljubimov |
|---|---|
| Předmět: |
Male
Adolescent Gene Expression Limbus Corneae Basement Membrane Adenoviridae stem cells cornea limbus Organ Culture Techniques Transduction Genetic Diabetes Mellitus Humans Child Eye Proteins Cells Cultured Aged Aged 80 and over Gene Expression Profiling Stem Cells Epithelium Corneal Genetic Therapy Middle Aged Proto-Oncogene Proteins c-met Immunohistochemistry eye diseases Female sense organs Autopsy Biomarkers Research Article |
| Zdroj: | ResearcherID Scopus-Elsevier Molecular Vision Europe PubMed Central |
| Popis: | Purpose We have previously identified specific epithelial proteins with altered expression in human diabetic central corneas. Decreased hepatocyte growth factor receptor (c-met) and increased proteinases were functionally implicated in the changes of these proteins in diabetes. The present study examined whether limbal stem cell marker patterns were altered in diabetic corneas and whether c-met gene overexpression could normalize these patterns. Methods Cryostat sections of 28 ex vivo and 26 organ-cultured autopsy human normal and diabetic corneas were examined by immunohistochemistry using antibodies to putative limbal stem cell markers including ATP-binding cassette sub-family G member 2 (ABCG2), N-cadherin, ΔNp63α, tenascin-C, laminin γ3 chain, keratins (K) K15, K17, K19, β1 integrin, vimentin, frizzled 7, and fibronectin. Organ-cultured diabetic corneas were studied upon transduction with adenovirus harboring c-met gene. Results Immunostaining for ABCG2, N-cadherin, ΔNp63α, K15, K17, K19, and β1 integrin, was significantly decreased in the stem cell-harboring diabetic limbal basal epithelium either by intensity or the number of positive cells. Basement membrane components, laminin γ3 chain, and fibronectin (but not tenascin-C) also showed a significant reduction in the ex vivo diabetic limbus. c-Met gene transduction, which normalizes diabetic marker expression and epithelial wound healing, was accompanied by increased limbal epithelial staining for K17, K19, ΔNp63α, and a diabetic marker α3β1 integrin, compared to vector-transduced corneas. Conclusions The data suggest that limbal stem cell compartment is altered in long-term diabetes. Gene therapy, such as with c-met overexpression, could be able to restore normal function to diabetic corneal epithelial stem cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|