Matthaei Mario, Zhu Angela Y, Kallay Laura, Eberhart Charles G, Cursiefen Claus, Jun Albert S
The Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, USA; Department of Ophthalmology, University of Cologne, Cologne, Germany; Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
The Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, USA.
Exp Eye Res. 2014 Dec;129:13-7. doi: 10.1016/j.exer.2014.10.011. Epub 2014 Oct 11.
Fuchs endothelial corneal dystrophy (FECD) is a genetically heterogeneous disease. Hypothesizing that cellular senescence may be relevant in FECD pathogenesis, genetically undifferentiated late-onset FECD endothelial samples were analyzed to identify common changes of specific senescence-related transcripts. Total RNA was extracted from 21 FECD endothelial samples retrieved from patients undergoing lamellar keratoplasty due to clinically diagnosed end-stage FECD and from 12 endothelial samples retrieved from normal autopsy eyes. Taqman low density array (TLDA) cards were used to analyze differential expression of 89 cellular senescence-related transcripts. Result validation was performed using individual real-time PCR assays. TLDA-analysis demonstrated differential expression of 31 transcripts (fold-change >1.5; p < 0.05). Thereof, 27 showed significant up-regulation and 4 significant down-regulation. Markedly elevated mRNA-levels of the constitutively active and reactive oxygen species-generating enzyme NOX4 were found in all evaluable FECD samples. In addition, increased expression of CDKN2A and its transcriptional activators ETS1 and ARHGAP18 (SENEX) along with decreased expression of CDKN2A inhibitor ID1 were detected in FECD samples. Consistent over-expression of NOX4 in FECD endothelial samples suggests a role as pathogenic factor and as a potential new treatment target in FECD. Transcriptional up-regulation of the CDKN2A-pathway provides further evidence for increased cellular senescence in FECD endothelium.
富克斯角膜内皮营养不良（FECD）是一种基因异质性疾病。假设细胞衰老可能与FECD发病机制相关，对基因未分化的迟发性FECD内皮样本进行分析，以确定特定衰老相关转录本的常见变化。从因临床诊断为终末期FECD而接受板层角膜移植术的患者的21个FECD内皮样本以及从正常尸检眼获取的12个内皮样本中提取总RNA。使用Taqman低密度阵列（TLDA）卡分析89个细胞衰老相关转录本的差异表达。使用个体实时PCR检测进行结果验证。TLDA分析显示31个转录本有差异表达（倍数变化>1.5；p<0.05）。其中，27个显示显著上调，4个显著下调。在所有可评估的FECD样本中均发现组成型活性和产生活性氧的酶NOX4的mRNA水平显著升高。此外，在FECD样本中检测到CDKN2A及其转录激活因子ETS1和ARHGAP18（SENEX）的表达增加，同时CDKN2A抑制剂ID1的表达降低。FECD内皮样本中NOX4的持续过表达表明其作为致病因素以及FECD潜在新治疗靶点的作用。CDKN2A通路的转录上调为FECD内皮细胞衰老增加提供了进一步证据。
