Investigating the Action and Physiological Role of Slc4a11 in the Cornea

研究 Slc4a11 在角膜中的作用和生理作用

基本信息

批准号：
10358498
负责人：
Mark Parker
金额：
$ 38.35万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-01 至 2023-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10358498
关键词：
Advanced Development Affect Air Altitude Animal Model Aqueous Humor Back Basement membrane Basic Science Bicarbonates Biomedical Engineering Blindness Cell model Cells Characteristics Chemicals Cicatrix Collagen Contact Lenses Cornea Corneal Diseases Corneal Endothelium Corneal Stroma Defect Development Disease Edema Endothelial Cells Endothelium Epithelial Eye Eyedrops Fluid Balance Genes Genetic Genetic Diseases Goals Health Human Humor Hydration status Immunohistochemistry Individual Inherited Ions Keratoplasty Kinetics Knockout Mice Knowledge Laboratories Light Link Liquid substance Long-Term Effects Longitudinal Studies Lubricants Membrane Membrane Transport Proteins Microelectrodes Mission Molecular Movement Mutation Operative Surgical Procedures Pharmacology Physiological Physiology Property Publications Pump Research Residual state Retina Role Sleep Structure Swelling Therapeutic Thinness Tissue Donors Transplantation United States Visual impairment Work Xenopus oocyte allograft rejection aqueous corneal allograft endothelial dysfunction gene therapy genetic linkage light scattering mathematical model novel novel strategies novel therapeutic intervention pH Homeostasis pediatric patients pressure programs repaired stem cells voltage clamp

项目摘要

Project Summary/Abstract Vision loss associated with corneal clouding affects ~300,000 individuals in the United States. The thickest layer of the cornea is the stroma; a transparent collagen matrix that tends to draw fluid from the aqueous humor. A layer of corneal endothelial cells between the stroma and aqueous humor provides little physical barrier to this fluid movement but actively extrudes osmolytes (bicarbonate lactate), and thus fluid, from the stroma back into the aqueous humor. Endothelial dysfunction allows fluid to accumulate in the stroma, distorting the matrix and causing it to scatter light. Corneal transplant has remained the definitive treatment for all corneal diseases for over a century, but advances our in our understanding of the genetics of disease and mechanisms of endothelial function have paved the way for development of bioengineered corneas and less invasive treatments. However, we still do not have a complete understanding of how the endothelium works. It is only relatively recently that genetic-linkage studies revealed the critical importance of the membrane transport protein SLC4A11 to endothelial health. We have recently established that SLC4A11 is a pH-sensitive H+ conductor. We hypothesize that SLC4A11 acts as a master regulator of endothelial cell pH, sensing and countering pH disturbance in the vicinity of pH-sensitive osmolyte transporters NBCe1-B (Na+/2HCO − cotransporter) and MCT1 (H+/lactate− 3 cotransporter). In the first part of our proposal we express SLC4A11 in Xenopus oocytes and, using a combination of ion-selective microelectrodes and voltage-clamp circuitry in order to determine the kinetic parameters governing SLC4A11 action that are necessary to implement SLC4A11 in mathematical models of endothelial fluid transport. We also examine the influence of SLC4A11 on NBCe1-B and MCT1 action. In the second part of our proposal we compare the progression of corneal disease signs in Slc4a11-knockout mice and our novel Nbce1b-knockout mouse to study the long-term effects of disturbed endothelial fluid transport. Finally, we investigate a novel therapeutic strategy to de-swell the edematous corneas of Slc4a11-knockout mice using eye drops. The long term goal of our research program is to understand how endothelial cells perform and balance fluid transport with pH homeostasis and to understand the importance of SLC4A11 to corneal health and disease. This project is aligned with the mission of the Corneal Disease Program of the NEI, to apply knowledge from basic science discoveries to the understanding of the physiology of the cornea and to the treatment of corneal disease.

项目概要/摘要在美国，与角膜混浊相关的视力丧失影响了大约 300,000 人。角膜的基质是透明的胶原基质，倾向于从房水 A 中吸取液体。基质和房水之间的角膜内皮细胞层对此几乎没有提供物理屏障液体运动，但主动将渗透剂（碳酸氢盐）和液体从基质挤出回房水功能障碍使液体积聚在基质中，扭曲基质并使其散射光的角膜移植仍然是所有角膜疾病的最终治疗方法。一个多世纪以来，我们对疾病遗传学和内皮细胞机制的理解取得了进展功能为生物工程角膜和微创治疗的发展铺平了道路。我们直到最近才完全了解内皮细胞的工作原理。遗传连锁研究揭示了膜转运蛋白 SLC4A11 对我们最近确定 SLC4A11 是一种 pH 敏感的 H+ 导体。 SLC4A11 充当内皮细胞 pH 值的主调节器，感知并对抗内皮细胞 pH 值的扰动 pH 敏感渗透剂转运蛋白 NBCe1-B（Na+/2HCO− 协同转运蛋白）和 MCT1（H+/乳酸−）附近 3 在我们提案的第一部分中，我们在非洲爪蟾卵母细胞中表达 SLC4A11，并使用离子选择性微电极和电压钳电路的组合，以确定动力学在数学模型中实现 SLC4A11 所必需的控制 SLC4A11 动作的参数我们还研究了 SLC4A11 对 NBCe1-B 和 MCT1 作用的影响。我们提案的第二部分我们比较了 Slc4a11 敲除小鼠的角膜疾病症状的进展情况我们的新型 Nbce1b 敲除小鼠用于研究内皮液体运输紊乱的长期影响。我们研究了一种新的治疗策略，使用 Slc4a11 敲除小鼠来消除水肿性角膜眼药水。我们研究计划的长期目标是了解内皮细胞如何发挥作用并平衡液体运输与 pH 稳态并了解 SLC4A11 对角膜健康和疾病的重要性。该项目与 NEI 角膜疾病项目的使命相一致，即应用来自了解角膜生理学和治疗角膜的基础科学发现疾病。