Centre for Genomic and Experimental Medicine, MRC Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, UK.
South East of Scotland Genetics Service, Western General Hospital, Edinburgh, UK.
Pediatr Pulmonol. 2024 Dec;59(12):3322-3332. doi: 10.1002/ppul.27200. Epub 2024 Aug 8.
Primary ciliary dyskinesia (PCD) is a genetic disorder affecting motile cilia. Most cases are inherited recessively, due to variants in >50 genes that result in abnormal or absent motile cilia. This leads to chronic upper and lower airway disease, subfertility, and laterality defects. Given overlapping clinical features and genetic heterogeneity, diagnosis can be difficult and often occurs late. Of those tested an estimated 30% of genetically screened PCD patients still lack a molecular diagnosis. A molecular diagnosis allows for appropriate clinical management including prediction of phenotypic features correlated to genotype. Here, we aimed to identify how readily a genetic diagnosis could be made using whole genome sequencing (WGS) to facilitate identification of pathogenic variants in known genes as well as novel PCD candidate genes.
WGS was used to screen for pathogenic variants in eight patients with PCD.
7/8 cases had homozygous or biallelic variants in DNAH5, DNAAF4 or DNAH11 classified as pathogenic or likely pathogenic. Three identified variants were deletions, ranging from 3 to 13 kb, for which WGS identified precise breakpoints, permitting confirmation by Sanger sequencing. WGS yielded identification of a de novo variant in a novel PCD gene TUBB4B.
Here, WGS uplifted genetic diagnosis of PCD by identifying structural variants and novel modes of inheritance in new candidate genes. WGS could be an important component of the PCD diagnostic toolkit, increasing molecular diagnostic yield from current (70%) levels, and enhancing our understanding of fundamental biology of motile cilia and variants in the noncoding genome.
原发性纤毛运动障碍(PCD)是一种影响游动纤毛的遗传疾病。大多数病例是由于 >50 个基因的变异导致异常或缺失的游动纤毛而隐性遗传的。这会导致慢性上呼吸道和下呼吸道疾病、不孕和偏侧性缺陷。由于具有重叠的临床特征和遗传异质性,诊断可能很困难,而且通常很晚才发生。在接受测试的人中,估计有 30%的基因筛查 PCD 患者仍然没有分子诊断。分子诊断可进行适当的临床管理,包括预测与基因型相关的表型特征。在这里,我们旨在确定使用全基因组测序(WGS)可以多容易地做出遗传诊断,以促进识别已知基因和新的 PCD 候选基因中的致病变异。
使用 WGS 筛选 8 例 PCD 患者的致病变异。
7/8 例 DNAH5、DNAAF4 或 DNAH11 中的纯合子或双等位基因变异被归类为致病性或可能致病性。鉴定出的三个变异是缺失,大小从 3 到 13kb,WGS 确定了精确的断点,允许通过 Sanger 测序进行确认。WGS 还鉴定出了一个新的 PCD 基因 TUBB4B 的新生变异。
在这里,WGS 通过鉴定新候选基因中的结构变异和新的遗传模式,提高了 PCD 的遗传诊断。WGS 可能成为 PCD 诊断工具包的重要组成部分,将目前(70%)的分子诊断率提高,并增强我们对运动纤毛和非编码基因组变异的基本生物学的理解。