Association of the Gutta-Induced Microenvironment With Corneal Endothelial Cell Behavior and Demise in Fuchs Endothelial Corneal Dystrophy
Association of the Gutta-Induced Microenvironment With Corneal Endothelial Cell Behavior and Demise in Fuchs Endothelial Corneal Dystrophy
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DOI：
10.1001/jamaophthalmol.2018.2031
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发表时间：
2018-08-01
影响因子：
8.1
通讯作者：
Jurkunas, Ula V.
中科院分区：
文献类型：
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作者：
Kocaba, Viridiana;Katildreddy, Kishore Reddy;Jurkunas, Ula V.
Fuchs内皮性角膜营养不良（FECD）患者肠管的数量和大小随着时间的推移而增加；然而，这些物理参数与疾病发病机制之间的关系尚不清楚。目的探讨绒毛在角膜内皮细胞功能中的作用。在体外模型中，来自人角膜内皮细胞系hccn - 21t的细胞被植入去细胞化的正常（n = 30）和FECD （n = 70）内皮基底（Descemet）膜（dm）上。正常的人角膜从三个来源送到我们的实验室。该研究于2015年9月至2017年7月在波士顿马萨诸塞州眼耳的Schepens眼科研究所进行。正常dm取自3个不同的组织库，fecd - dm取自2个科室接受内皮角膜移植术的患者。内皮细胞的形状、生长和迁移通过活细胞成像进行评估，基因表达分析作为导管直径的函数通过激光捕获显微镜进行评估。结果正常dms供者的平均（SD）年龄为65.6（4.4）岁（16名女性[53%]），FECD-DMs供者的平均（SD）年龄为68.9（10.6）岁（43名女性[61%]）。与FECD dm相比，正常dm上细胞覆盖面积更大（平均[SD]， 97.7%[8.5%]），平均（SD）细胞数（2083[153]个细胞/mm(2)）更大（72.8% [11%],P = 0.02和1541[221]个细胞/mm(2)） 221/mm(2)；P = 0.01)。在FECD DMs上观察到内皮细胞生长在肠管上的差异，这取决于肠管直径。平均直径为10.5 (2.9)μ m的Guttae不影响细胞生长，而直径为21.1(4.9)μ m的Guttae仅被细胞质覆盖。最大平均直径（SD）为31.8 (3.8)mu m的Guttae没有被细胞覆盖，而是被细胞包围成玫瑰花状。此外，与生长在正常dm或小肠上的细胞相比，大肠旁的细胞上调了aSMA、N-cadherin、Snail1和NOX4基因。此外，与离体FECD标本类似，大导管诱导了玫瑰花状的tunel阳性细胞凋亡。结论和相关性这些研究结果强调了guttae在内皮细胞生长、迁移和存活中的重要作用。这些数据表明，如果随后的临床研究证实了这些实验室发现，FECD的细胞治疗程序可能会以宿主肠管的直径为指导。
IMPORTANCE The number and size of guttae increase over time in Fuchs endothelial corneal dystrophy (FECD); however, the association between these physical parameters and disease pathogenesis is unclear.OBJECTIVE To determine the role of guttae in corneal endothelial cell function.DESIGN, SETTINGS, AND PARTICIPANTS In an in vitro model, cells from a human corneal endothelial cell line, HCENC-21T, were seeded on decellularized normal (n = 30) and FECD (n = 70) endothelial basement (Descemet) membranes (DMs). Normal human corneas were sent to our laboratory from 3 sources. The study took place at the Schepens Eye Research Institute, Massachusetts Eye and Ear, Boston, and was performed from September 2015 to July 2017. Normal DMs were obtained from 3 different tissue banks and FECD-DMs were obtained from patients undergoing endothelial keratoplasty in 2 departments.MAIN OUTCOMES AND MEASURES Endothelial cell shape, growth, and migration were assessed by live-cell imaging, and gene expression analysis as a function of guttae diameter was assessed by laser capture microscopy.RESULTS Mean (SD) age of normal-DMs donors was 65.6 (4.4) years (16 women [53%]), and mean (SD) age of FECD-DMs donors was 68.9 (10.6) years (43 women [61%]). Cells covered a greater area (mean [SD], 97.7% [8.5%]) with a greater mean (SD) number of cells (2083 [153] cells/mm(2)) on the normal DMs compared with the FECD DMs (72.8% [11%]; P = .02 and 1541 [221] cells/mm(2)221/mm(2); P = .01, respectively). Differences in endothelial cell growth over guttae were observed on FECD DMs depending on the guttae diameter. Guttae with a mean (SD) diameter of 10.5 (2.9) mu m did not impede cell growth, whereas those with a diameter of 21.1(4.9) mu m were covered only by the cell cytoplasm. Guttae with the largest mean (SD) diameter, 31.8 (3.8) mu m, were not covered by cells, which instead surrounded them in a rosette pattern. Moreover, cells adjacent to large guttae upregulated aSMA, N-cadherin, Snail1, and NOX4 genes compared with ones grown on normal DMs or small guttae. Furthermore, large guttae induced TUNEL-positive apoptosis in a rosette pattern, similar to ex vivo FECD specimens.CONCLUSIONS AND RELEVANCE These findings highlight the important role of guttae in endothelial cell growth, migration, and survival. These data suggest that cell therapy procedures in FECD might be guided by the diameter of the host guttae if subsequent clinical studies confirm these laboratory findings.