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

• 批准号: 10090471
• 负责人: Mark Parker
• 金额: $ 38.36万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10090471
• 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

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.

项目摘要/摘要 与角膜云相关的视力损失会影响美国约30万人。最厚的层 角膜是基质；一种透明的胶原蛋白基质，倾向于从水性幽默中吸收流体。一个 基质和水性幽默之间的角膜内皮细胞层几乎没有物理障碍 流体运动，但积极地挤压渗透液（碳酸氢盐缝合），从而从基质中回到 水性幽默。内皮功能障碍允许液体积聚在基质中，使基质扭曲和 使其散布光。角膜移植仍然是所有角膜疾病的明确治疗方法 一个多世纪以来，我们在对疾病遗传学和内皮机制的理解方面促进了我们的发展 功能为开发生物工程角膜和侵入性较少的疗法铺平了道路。然而， 我们仍然对内皮的工作原理不完全了解。直到最近 遗传链接研究揭示了膜转运蛋白SLC4A11至关重要的重要性 内皮健康。我们最近确定SLC4A11是pH敏感的H+导体。我们假设 SLC4A11充当内皮细胞pH的主要调节器，感知和反击pH灾难 pH敏感性渗透转运蛋白的附近NBCE1-B（Na+/2HCO-共转运蛋白）和MCT1（H+/乳酸 - 3 共转运者）。在提案的第一部分中，我们在Xenopus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 管理SLC4A11操作的参数是在数学模型中实现SLC4A11所必需的 内皮流体转运。我们还检查了SLC4A11对NBCE1-B和MCT1作用的影响。在 提案的第二部分我们比较了SLC4A11敲除小鼠中角膜疾病迹象的进展 我们的新型NBCE1b-KNOCKOUT小鼠研究了干扰内皮流体转运的长期影响。最后， 我们研究了一种新的理论策略，以使用SLC4A11-敲除小鼠的水性角膜进行脱水 眼睛滴。 我们的研究计划的长期目标是了解内皮细胞的性能和平衡流体 具有pH稳态的运输，并了解SLC4A11对角膜健康和疾病的重要性。 该项目与NEI角膜疾病计划的使命保持一致，以应用 基础科学发现，了解角膜生理学和角膜的治疗 疾病。