Molecular and functional characterization of the regenerative potential of slow cycling corneal epithelial cells

描述(由申请人提供)：角膜缘干细胞缺乏症(LSCD)的特征是眼痛、再上皮化不良、角膜溃疡、结膜细胞内生和视力丧失。LSCD可能继发于多种疾病，如Steven Johnson综合征、眼瘢痕性类天疱疮、慢性炎症和化学损伤。将角膜上皮干细胞移植到干细胞缺陷性角膜有可能使盲眼恢复视力。改善干细胞缺乏症患者的康复需要更好地利用和诱导成人角膜上皮干细胞的再生能力。这项拟议的研究将利用干细胞的慢循环表型来帮助识别、纯化和扩增角膜干细胞群体，使用小鼠转基因“脉冲追逐系统”。目的1研究小鼠角膜中慢周期细胞的基因表达谱。该系统将在SubAim 1.1中进行优化，分离的慢周期细胞将在SubAim 1.2中进行基因表达分析。目的2将使用体外和体内模型来评估慢周期角膜上皮细胞的再生潜力。最后，Aim 3将确定慢周期细胞相关基因，这些基因对角膜上皮再生至关重要。

DESCRIPTION (provided by applicant): Limbal Stem Cell Deficiency (LSCD) is characterized by ocular pain, poor re-epithelialization, corneal ulceration, conjunctival cell ingrowth and loss f visual acuity. LSCD may be secondary to myriad conditions such as Steven Johnson's Syndrome, ocular cicatricial pemphigoid, chronic inflammation and chemical injury. Transplantation of corneal epithelial stem cells to stem cell deficient corneas has the potential t restore sight to blind eyes. Improving the rehabilitation of patients with stem cell deficiency wil require the ability to better harness and induce the regenerative capacity of adult corneal epithelial stem cells. The proposed research will capitalize upon the slow cycling phenotype of stem cells to aid in identifying, purifying and expanding corneal stem epithelial cell populations using a murine transgenic "pulse-chase system". Aim 1 will characterize the gene expression profile of slow cycling cells in the murine cornea. The system will be optimized in subaim 1.1 and isolated slow cycling cells will undergo gene expression analysis in subaim 1.2. Aim 2 will use in vitro and in vivo models to evaluate the regenerative potential of slow cycling corneal epithelial cells. Finally, Aim 3 will identify slow cycling cell related genes that are critical fo corneal epithelium regeneration.