INSERM/UMR S933, Université Pierre et Marie Curie-Paris 6, Paris, France.
Am J Hum Genet. 2013 Sep 5;93(3):561-70. doi: 10.1016/j.ajhg.2013.07.013. Epub 2013 Aug 29.
Primary ciliary dyskinesia (PCD) is a rare autosomal-recessive respiratory disorder resulting from defects of motile cilia. Various axonemal ultrastructural phenotypes have been observed, including one with so-called central-complex (CC) defects, whose molecular basis remains unexplained in most cases. To identify genes involved in this phenotype, whose diagnosis can be particularly difficult to establish, we combined homozygosity mapping and whole-exome sequencing in a consanguineous individual with CC defects. This identified a nonsense mutation in RSPH1, a gene whose ortholog in Chlamydomonas reinhardtii encodes a radial-spoke (RS)-head protein and is mainly expressed in respiratory and testis cells. Subsequent analyses of RSPH1 identified biallelic mutations in 10 of 48 independent families affected by CC defects. These mutations include splicing defects, as demonstrated by the study of RSPH1 transcripts obtained from airway cells of affected individuals. Wild-type RSPH1 localizes within cilia of airway cells, but we were unable to detect it in an individual with RSPH1 loss-of-function mutations. High-speed-videomicroscopy analyses revealed the coexistence of different ciliary beating patterns-cilia with a normal beat frequency but abnormal motion alongside immotile cilia or cilia with a slowed beat frequency-in each individual. This study shows that this gene is mutated in 20.8% of individuals with CC defects, whose diagnosis could now be improved by molecular screening. RSPH1 mutations thus appear as a major etiology for this PCD phenotype, which in fact includes RS defects, thereby unveiling the importance of RSPH1 in the proper building of CCs and RSs in humans.
原发性纤毛运动障碍(PCD)是一种罕见的常染色体隐性呼吸道疾病,由运动纤毛的缺陷引起。已经观察到各种轴丝超微结构表型,包括一种所谓的中央复合物(CC)缺陷,在大多数情况下,其分子基础仍未得到解释。为了确定与这种表型相关的基因,因为这种表型的诊断特别难以建立,我们在一个具有 CC 缺陷的近亲个体中结合了纯合子作图和全外显子组测序。这在 RSPH1 基因中发现了一个无义突变,该基因在 Chlamydomonas reinhardtii 中的同源物编码一个放射辐(RS)头蛋白,主要在呼吸和睾丸细胞中表达。随后对 RSPH1 的分析在 48 个受 CC 缺陷影响的独立家族中的 10 个中发现了双等位基因突变。这些突变包括剪接缺陷,这是通过研究受影响个体气道细胞中获得的 RSPH1 转录本来证明的。野生型 RSPH1 定位于气道细胞的纤毛内,但我们无法在具有 RSPH1 功能丧失突变的个体中检测到它。高速视频显微镜分析显示,在每个个体中,不同的纤毛跳动模式共存 - 具有正常跳动频率但运动异常的纤毛与不动纤毛或跳动频率减慢的纤毛并存。这项研究表明,该基因在 20.8%的 CC 缺陷个体中发生突变,现在可以通过分子筛查来改善这些个体的诊断。RSPH1 突变因此成为这种 PCD 表型的主要病因,实际上包括 RS 缺陷,从而揭示了 RSPH1 在人类 CC 和 RS 正确构建中的重要性。