Alterations of epithelial stem cell marker patterns in human diabetic corneas and effects of c-met gene therapy

作者：Saghizadeh Mehrnoosh; Soleymani Siavash; Harounian Angel; Bhakta Bhavik; Troyanovsky Sergey M; Brunken William J; Pellegrini Graziella; Ljubimov Alexander V^*

来源：Molecular Vision, 2011, 17(235-36): 2177-2190.

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, Delta Np63 alpha, tenascin-C, laminin gamma 3 chain, keratins (K) K15, K17, K19, beta(1) 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, Delta Np63 alpha, K15, K17, K19, and beta(1) 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 gamma 3 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, Delta Np63 alpha, and a diabetic marker alpha(3)beta(1) 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.

• 出版日期2011-8-12
• 单位西北大学