Corneal endothelial cell survival in human donor corneas for transplantation

用于移植的人供体角膜中角膜内皮细胞的存活率

• 批准号: 8204537
• 负责人: SHAY-WHEY M KOH
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204537
• 关键词: Amino Acids; Apoptosis; Apoptotic; Cell Adhesion Molecules; Cell Density; Cell Differentiation process; Cell Size; Cell Survival; Cell physiology; Cell-Cell Adhesion; Ciliary Neurotrophic Factor; Ciliary Neurotrophic Factor Receptor; Cornea; Corneal Endothelium; Descemet' s membrane; Development; Differentiation Antigens; Endothelial Cells; Eye; Eye Banks; Failure; Future; Goals; Human; In Situ; In Vitro; Injury; Keratoplasty; Knowledge; Life; Maintenance; Modeling; N-Cadherin; Operating Rooms; Oxidative Stress; Physiology; Procedures; Proteins; Reporting; Risk Factors; Shapes; Techniques; Thick; Time; Transplantation; Vasoactive Intestinal Peptide; abstracting; anterior chamber; autocrine; graft failure; killings; neurotrophic factor; release factor

Abstract Corneal endothelial (CE) cell loss is a critical risk factor in corneal graft failure at any time in the life of the graft, which can be as late as 5-10 years after an initially successful transplant. A new procedure, Descemet's stripping automated endothelial keratoplasty (DSAEK), which is superior to the traditional technique in many aspects, requires extensive manipulation of the corneal endothelium, resulting in extensive CE cell loss. Thus, future development of late CE failure in eyes that have received DSAEK is likely. Previously, We have reported how a CE autocrine trophic factor, the 28 amino-acid vasoactive intestinal peptide (VIP) maintains the differentiated state of the corneal endothelium, including CE cell size, shape, and retention, in situ in the human donor cornea, whereas exogenous VIP protects it against the killing effect of oxidative stress. VIP increases synthesis of the CE cell differentiation marker N-cadherin, the cell-cell adhesion molecule, and that of the anti-apoptotic protein Bcl-2. The endogenous VIP in CE cells is upregulated by ciliary neurotrophic factor (CNTF), which is also a CE autocrine trophic factor released by CE cells surviving the oxidative stress. The CNTF receptor (CNTFR¿) is expressed in CE cells in human donor cornea and gradually becomes lost during corneal storage. By applying this knowledge gained from studying CE cell physiology to the enhancement of CE cell survival in corneal transplantation, which is our goal, we will also validate the relevance of this knowledge. Our specific aims are to demonstrate that 1. VIP treatment prior to storage of freshly dissected human donor corneas increases their CE cell N-cadherin, Bcl-2, CNTFR¿, and retention levels in corneal storage, 2. One additional VIP treatment at the time of microkeratome cutting of the human donor corneas stored for DSAEK brings about beneficial effects described in aim 1 and decreased CE cell apoptosis and injury to precut corneas for DSAEK, 3. VIP treatments prior to storage of freshly dissected human donor corneas and prior to microkeratome cutting produce superior precut corneas for DSAEK demonstrating decreased CE damage in an established in vitro endothelial keratoplasty model, 4. Intra-operative VIP (or CNTF) treatment decreases CE damage in an established in vitro endothelial keratoplasty model.

摘要 角膜内皮（CE）细胞丢失是任何情况下角膜移植失败的关键风险因素。 在移植物的生命周期中的时间，这可能是在最初成功的移植后5-10年。 移植后弹力层剥脱式自动角膜内皮移植术 DSAEK技术在许多方面上级传统技术， 角膜内皮的广泛操作，导致大量CE细胞损失。 因此，在接受DSAEK的眼睛中，未来可能会发生晚期CE失败。 以前，我们已经报道了如何CE自分泌营养因子，28氨基酸 血管活性肠肽（VIP）维持角膜的分化状态， 内皮，包括CE细胞大小、形状和保留，在人供体中原位 角膜，而外源性VIP保护其免受氧化应激的杀伤作用。 VIP增加CE细胞分化标记物N-钙粘蛋白的合成， 粘附分子，以及抗凋亡蛋白Bcl-2。内源性VIP CE细胞被睫状神经营养因子（CNTF）上调，睫状神经营养因子也是一种CE细胞。 自分泌营养因子由在氧化应激中存活的CE细胞释放。CNTF 受体（CNTFR）在人供体角膜的CE细胞中表达，并逐渐 在角膜储存期间丢失。通过应用这些从学习中获得的知识， CE细胞生理学对角膜移植中CE细胞存活的增强， 这是我们的目标，我们也将验证这些知识的相关性。我们的具体 目的是证明， 1.新鲜解剖的人类供体角膜储存前的VIP处理增加 他们的CE细胞N-钙粘蛋白，Bcl-2，CNTFR和角膜储存中的保留水平， 2.在微型角膜刀切割人体时进行一次额外VIP治疗 为DSAEK储存的供体角膜带来了目标中描述的有益效果 1和减少CE细胞凋亡和DSAEK对预切角膜的损伤， 3.新鲜解剖的人类供体角膜储存前的VIP处理， 在微型角膜刀切割之前，为DSAEK生产上级的预切角膜 证明在已建立的体外内皮细胞中CE损伤降低， 角膜移植模型， 4.术中VIP（或CNTF）治疗可减少已建立的CE损伤。 体外内皮角膜移植模型。