Le Derek J, Chung Duk-Won D, Frausto Ricardo F, Kim Michelle J, Aldave Anthony J
Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, California, United States of America.
PLoS One. 2016 Jun 29;11(6):e0158467. doi: 10.1371/journal.pone.0158467. eCollection 2016.
Posterior polymorphous corneal dystrophy 1 (PPCD1) is a genetic disorder that affects corneal endothelial cell function and leads to loss of visual acuity. PPCD1 has been linked to a locus on chromosome 20 in multiple families; however, Sanger sequencing of protein-coding genes in the consensus region failed to identify any causative missense mutations. In this study, custom capture probes were utilized for targeted next-generation sequencing of the linked region in a previously reported family with PPCD1. Variants were detected through two bioinformatics pipelines and filtered according to multiple criteria. Additionally, a high-resolution microarray was used to detect copy number variations. No non-synonymous variants in the protein-coding region of annotated genes were identified. However, 12 single nucleotide variants in 10 genes, and 9 indels in 7 genes met the filtering criteria and were considered candidate variants for PPCD1. Eleven single nucleotide variants were confirmed by Sanger sequencing, including 2 synonymous variants and 9 non-coding variants, in 9 genes. One microdeletion was detected in an intron of OVOL2 by microarray but was subsequently not identified by PCR. Using a comprehensive next-generation sequencing approach, a total of 16 genes containing single nucleotide variants or indels that segregated with the affected phenotype in an affected family previously mapped to the PPCD1 locus were identified. Screening of these candidate genes in other families previously mapped to the PPCD1 locus will likely result in the identification of the genetic basis of PPCD1.
后极性多形性角膜营养不良1型(PPCD1)是一种影响角膜内皮细胞功能并导致视力丧失的遗传性疾病。PPCD1在多个家族中与20号染色体上的一个位点相关联;然而,对共有区域中蛋白质编码基因进行Sanger测序未能鉴定出任何致病性错义突变。在本研究中,定制捕获探针用于对先前报道的一个患有PPCD1的家族中相关区域进行靶向二代测序。通过两种生物信息学流程检测变异,并根据多种标准进行筛选。此外,使用高分辨率微阵列检测拷贝数变异。在注释基因的蛋白质编码区域未鉴定出非同义变异。然而,10个基因中的12个单核苷酸变异以及7个基因中的9个插入/缺失符合筛选标准,被视为PPCD1的候选变异。通过Sanger测序确认了9个基因中的11个单核苷酸变异,包括2个同义变异和9个非编码变异。通过微阵列在OVOL2的一个内含子中检测到一个微缺失,但随后通过PCR未鉴定到。使用全面的二代测序方法,在一个先前定位到PPCD1位点的患病家族中,共鉴定出16个含有与患病表型共分离的单核苷酸变异或插入/缺失的基因。在其他先前定位到PPCD1位点的家族中对这些候选基因进行筛选,可能会确定PPCD1的遗传基础。