Centre de Recherche du CHU de Québec-Université Laval, Hôpital du Saint-Sacrement, Québec, Québec, Canada; Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/Laboratoire d'Organogenèse Experimental, Québec, Québec, Canada.
Maisonneuve-Rosemont Hospital Research Center, Montreal, Québec, Canada; Department of Ophthalmology, University of Montreal, Montreal, Québec, Canada.
Am J Pathol. 2018 Jul;188(7):1703-1712. doi: 10.1016/j.ajpath.2018.03.014. Epub 2018 Apr 24.
Fuchs endothelial corneal dystrophy (FECD) is a corneal pathology that affects the endothelial cell's ability to maintain deturgescence, resulting in a progressive loss of corneal transparency. In this study, we investigated the expression of function-related proteins in corneal endothelial cells using FECD or healthy corneal endothelial cells, either in a cell culture two-dimensional model or in an engineered corneal endothelium three-dimensional tissue model. No statistically significant difference in gene regulation was observed for the function-related families ATP1, SLC4, SLC16, AQP, TJP, and CDH between the FECD and the healthy cell models. Similarly, no difference in barrier integrity (transendothelial electrical resistance measurements and permeability assays) was observed in vitro between FECD and healthy cultured cells. Protein expression of the key function-related families was decreased for Na/K-ATPase α1 subunit, monocarboxylate transporters 1 and 4 in native ex vivo end-stage FECD specimens, whereas it returned to levels comparable to that of healthy tissues in the engineered FECD model. These results indicate that cell expansion and tissue engineering culture conditions can generate a corneal endothelium from pathologic FECD cells, with levels of function-related proteins similar to that of healthy tissues. Overall, these results explain why it is possible to reform a functional endothelium using corneal endothelial cells isolated from nonfunctional FECD pathologic specimens.
Fuchs 内皮角膜营养不良（FECD）是一种角膜病理，它影响内皮细胞维持去水肿的能力，导致角膜透明度逐渐丧失。在这项研究中，我们使用 FECD 或健康的角膜内皮细胞，在细胞培养二维模型或工程化角膜内皮三维组织模型中，研究了角膜内皮细胞中功能相关蛋白的表达。在功能相关家族 ATP1、SLC4、SLC16、AQP、TJP 和 CDH 的基因调控方面，FECD 和健康细胞模型之间没有观察到统计学上的显著差异。同样，在体外，FECD 和健康培养细胞之间的屏障完整性（跨内皮电阻测量和通透性测定）也没有差异。在天然的 FECD 终末期标本中，关键功能相关家族的蛋白表达减少，包括 Na/K-ATPase α1 亚基、单羧酸转运蛋白 1 和 4，而在工程化的 FECD 模型中，其表达水平恢复到与健康组织相当的水平。这些结果表明，细胞扩增和组织工程培养条件可以从病变的 FECD 细胞中产生具有类似健康组织功能相关蛋白水平的角膜内皮。总的来说，这些结果解释了为什么可以使用从非功能性 FECD 病变标本中分离出的角膜内皮细胞来重建功能性内皮。
