The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Am J Hum Genet. 2012 Mar 9;90(3):533-9. doi: 10.1016/j.ajhg.2012.01.013. Epub 2012 Feb 16.
Fuchs corneal dystrophy (FCD) is a genetic disorder of the corneal endothelium and is the most common cause of corneal transplantation in the United States. Previously, we mapped a late-onset FCD locus, FCD2, on chromosome 18q. Here, we present next-generation sequencing of all coding exons in the FCD2 critical interval in a multigenerational pedigree in which FCD segregates as an autosomal-dominant trait. We identified a missense change in LOXHD1, a gene causing progressive hearing loss in humans, as the sole variant capable of explaining the phenotype in this pedigree. We observed LOXHD1 mRNA in cultured human corneal endothelial cells, whereas antibody staining of both human and mouse corneas showed staining in the corneal epithelium and endothelium. Corneal sections of the original proband were stained for LOXHD1 and demonstrated a distinct increase in antibody punctate staining in the endothelium and Descemet membrane; punctate staining was absent from both normal corneas and FCD corneas negative for causal LOXHD1 mutations. Subsequent interrogation of a cohort of >200 sporadic affected individuals identified another 15 heterozygous missense mutations that were absent from >800 control chromosomes. Furthermore, in silico analyses predicted that these mutations reside on the surface of the protein and are likely to affect the protein's interface and protein-protein interactions. Finally, expression of the familial LOXHD1 mutant allele as well as two sporadic mutations in cells revealed prominent cytoplasmic aggregates reminiscent of the corneal phenotype. All together, our data implicate rare alleles in LOXHD1 in the pathogenesis of FCD and highlight how different mutations in the same locus can potentially produce diverse phenotypes.
Fuchs 角膜营养不良(FCD)是一种角膜内皮的遗传性疾病,是美国最常见的角膜移植原因。先前,我们在染色体 18q 上定位了一个迟发性 FCD 基因座 FCD2。在此,我们对一个多代家族中 FCD2 关键区间的所有编码外显子进行了下一代测序,该家族中 FCD 作为常染色体显性遗传。我们在该家系中发现了 LOXHD1 基因的错义改变,该基因导致人类进行性听力损失,是唯一能够解释该家系表型的变异。我们在培养的人角膜内皮细胞中观察到 LOXHD1mRNA,而人角膜和小鼠角膜的抗体染色显示在角膜上皮和内皮中有染色。原始先证者的角膜切片用 LOXHD1 染色,在内皮和 Descemet 膜中显示出抗体点状染色明显增加;正常角膜和无因果 LOXHD1 突变的 FCD 角膜均无点状染色。随后对>200 例散发性受累个体的队列进行了进一步检测,发现了另外 15 个杂合错义突变,这些突变在>800 个对照染色体中均不存在。此外,计算机模拟分析预测这些突变位于蛋白质表面,可能影响蛋白质的界面和蛋白-蛋白相互作用。最后,在细胞中表达家族性 LOXHD1 突变等位基因和两个散发性突变显示出明显的细胞质聚集,类似于角膜表型。总之,我们的数据表明 LOXHD1 的罕见等位基因参与了 FCD 的发病机制,并强调了同一基因座中的不同突变如何可能产生不同的表型。