Department of Human Genetics, Institute of Ophthalmology, UCL, London, UK.
Moorfields Eye Hospital, London, UK.
Hum Mutat. 2013 Nov;34(11):1537-1546. doi: 10.1002/humu.22398. Epub 2013 Sep 17.
This study was undertaken to investigate the prevalence of sequence variants in LCA5 in patients with Leber congenital amaurosis (LCA), early-onset retinal dystrophy (EORD), and autosomal recessive retinitis pigmentosa (arRP); to delineate the ocular phenotypes; and to provide an overview of all published LCA5 variants in an online database. Patients underwent standard ophthalmic evaluations after providing informed consent. In selected patients, optical coherence tomography (OCT) and fundus autofluorescence imaging were possible. DNA samples from 797 unrelated patients with LCA and 211 with the various types of retinitis pigmentosa (RP) were screened by Sanger sequence analysis of all LCA5 exons and intron/exon junctions. Some LCA patients were prescreened by APEX technology or selected based on homozygosity mapping. In silico analyses were performed to assess the pathogenicity of the variants. Segregation analysis was performed where possible. Published and novel LCA5 variants were collected, amended for their correct nomenclature, and listed in a Leiden Open Variation Database (LOVD). Sequence analysis identified 18 new probands with 19 different LCA5 variants. Seventeen of the 19 LCA5 variants were novel. Except for two missense variants and one splice site variant, all variants were protein-truncating mutations. Most patients expressed a severe phenotype, typical of LCA. However, some LCA subjects had better vision and intact inner segment/outer segment (IS/OS) junctions on OCT imaging. In two families with LCA5 variants, the phenotype was more compatible with EORD with affected individuals displaying preserved islands of retinal pigment epithelium. One of the families with a milder phenotype harbored a homozygous splice site mutation; a second family was found to have a combination of a stop mutation and a missense mutation. This is the largest LCA5 study to date. We sequenced 1,008 patients (797 with LCA, 211 with arRP) and identified 18 probands with LCA5 mutations. Mutations in LCA5 are a rare cause of childhood retinal dystrophy accounting for ∼2% of disease in this cohort, and the majority of LCA5 mutations are likely null. The LCA5 protein truncating mutations are predominantly associated with LCA. However, in two families with the milder EORD, the LCA5 gene analysis revealed a homozygous splice site mutation in one and a stop mutation in combination with a missense mutation in a second family, suggesting that this milder phenotype is due to residual function of lebercilin and expanding the currently known phenotypic spectrum to include the milder early onset RP. Some patients have remaining foveal cone structures (intact IS/OS junctions on OCT imaging) and remaining visual acuities, which may bode well for upcoming treatment trials.
本研究旨在调查 LCA5 序列变异在莱伯先天性黑矇(LCA)、早发性视网膜营养不良(EORD)和常染色体隐性视网膜色素变性(arRP)患者中的发生率;描述眼部表型;并在在线数据库中提供所有已发表的 LCA5 变异的概述。患者在提供知情同意后接受了标准的眼科评估。在选定的患者中,可以进行光学相干断层扫描(OCT)和眼底自发荧光成像。通过对所有 LCA5 外显子和内含子/外显子接头的 Sanger 序列分析,对 797 名无血缘关系的 LCA 患者和 211 名各种类型的视网膜色素变性(RP)患者的 DNA 样本进行了筛查。一些 LCA 患者通过 APEX 技术进行了预筛查,或根据纯合性作图进行了选择。对变异进行了体外分析以评估其致病性。在可能的情况下进行了分离分析。收集并修正了已发表和新的 LCA5 变异,将其列入莱顿开放变异数据库(LOVD)。序列分析确定了 18 名新的先证者,有 19 种不同的 LCA5 变异。19 种 LCA5 变异中有 17 种是新的。除了两个错义变异和一个剪接位点变异外,所有变异均为蛋白截断突变。大多数患者表现出严重的表型,与 LCA 典型表型一致。然而,一些 LCA 患者的 OCT 成像上具有更好的视力和完整的内节/外节(IS/OS)连接。在两个具有 LCA5 变异的家庭中,表型与 EORD 更吻合,受影响的个体表现出保留的视网膜色素上皮岛。具有更温和表型的一个家庭携带有纯合剪接位点突变;第二个家庭被发现存在一个终止突变和一个错义突变的组合。这是迄今为止最大的 LCA5 研究。我们对 1008 名患者(797 名 LCA,211 名 arRP)进行了测序,发现了 18 名携带 LCA5 突变的先证者。LCA5 突变是儿童视网膜营养不良的罕见原因,占该队列疾病的约 2%,大多数 LCA5 突变很可能是无效的。LCA5 蛋白截断突变主要与 LCA 相关。然而,在两个具有更温和 EORD 的家庭中,LCA5 基因分析显示一个家庭存在纯合剪接位点突变,另一个家庭存在终止突变和错义突变的组合,表明这种更温和的表型是由于 lebercilin 的残留功能所致,并将目前已知的表型谱扩展到包括更温和的早发性 RP。一些患者仍具有完整的中心凹锥细胞结构(OCT 成像上的完整 IS/OS 连接)和残余视力,这可能为即将进行的治疗试验带来良好的效果。