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

慢循环角膜上皮细胞再生潜力的分子和功能表征

• 批准号: 9087268
• 负责人: Mark I Rosenblatt
• 金额: $ 19.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9087268
• 关键词: Adult; Biological Assay; Blindness; Candidate Disease Gene; Cell Cycle; Cell Nucleus; Cell Separation; Cells; Chemical Injury; Chronic; Complementary DNA; Cornea; Corneal Ulcer; Data; Defect; Doxycycline; Epithelial; Epithelial Cells; Excision; Eye; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Health; Image; In Vitro; Infection; Inflammation; Label; Lead; Lentivirus Vector; Limbus Corneae; Mechanics; Modeling; Molecular; Molecular Analysis; Molecular Profiling; Mus; Natural regeneration; Nuclear; Ocular cicatricial pemphigoid; Patients; Phenotype; Physiologic pulse; Population; Production; Rehabilitation therapy; Research; Risk; Secondary to; Skin; Stem cells; Stevens-Johnson Syndrome; System; Time; Tissues; Transgenic Organisms; Transplantation; Trauma; Vision; Visual Acuity; Work; blind; corneal epithelial stem cells; corneal epithelium; differential expression; enhanced green fluorescent protein; improved; in vivo; in vivo Model; knock-down; limbal; novel; novel therapeutics; ocular pain; ocular surface; overexpression; regenerative; slow potential; small hairpin RNA; stem; stemness; transcriptome sequencing

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.

描述（由申请人提供）：角膜缘干细胞缺乏症（LSCD）的特征是眼部疼痛、上皮再生不良、角膜溃疡、结膜细胞向内生长和视力丧失。LSCD可能继发于多种疾病，如史蒂文约翰逊综合征、眼瘢痕性类天疱疮、慢性炎症和化学损伤。角膜上皮干细胞移植到干细胞缺陷的角膜中具有使盲眼恢复视力的潜力。改善干细胞缺陷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。拟议的研究将利用干细胞的慢循环表型，以帮助识别，纯化和扩大角膜干上皮细胞群体使用鼠转基因“脉冲追逐系统”。目的1研究小鼠角膜慢循环细胞的基因表达谱。将在子目标1.1中优化系统，并在子目标1.2中对分离的慢循环细胞进行基因表达分析。目的2将利用体外和体内模型来评价慢循环角膜上皮细胞的再生潜力。最后，目标3将鉴定对角膜上皮再生至关重要的慢循环细胞相关基因。