PDF(Pubmed)
Abstract：
Primary ciliary dyskinesia (PCD) is a rare disorder characterized by extensive genetic heterogeneity. However, in the genetic pathogenesis of PCD, copy number variation (CNV) has not received sufficient attention and has rarely been reported, especially in China. Next-generation sequencing (NGS) followed by targeted CNV analysis was used in patients highly suspected to have PCD with negative results in routine whole-exome sequencing (WES) analysis. Quantitative real-time polymerase chain reaction (qPCR) and Sanger sequencing were used to confirm these CNVs. To further characterize the ciliary phenotypes, high-speed video microscopy analysis (HSVA), transmission electron microscopy (TEM), and immunofluorescence (IF) analysis were used. Patient 1 (F1: II-1), a 0.6-year-old girl, came from a nonconsanguineous family-I. She presented with situs inversus totalis, neonatal respiratory distress, and sinusitis. The nasal nitric oxide level was markedly reduced. The respiratory cilia beat with reduced amplitude. TEM revealed shortened outer dynein arms (ODA) of cilia. chr5:13717907-13722661del spanning exons 71-72 was identified by NGS-based CNV analysis. Patient 2 (F2: IV-4), a 37-year-old man, and his eldest brother Patient 3 (F2: IV-2) came from a consanguineous family-II. Both had sinusitis, bronchiectasis and situs inversus totalis. The respiratory cilia of Patient 2 and Patient 3 were found to be uniformly immotile, with ODA defects. Two novel homozygous deletions chr5:13720087_13733030delinsGTTTTC and chr5:13649539_1 3707643del, spanning exons 69-71 and exons 77-79 were identified by NGS-based CNV analysis. Abnormalities in DNA copy number were confirmed by qPCR amplification. IF showed that the respiratory cilia of Patient 1 and Patient 2 were deficient in dynein axonemal heavy chain 5 (DNAH5) protein expression. This report identified three novel DNAH5 disease-associated variants by WES-based CNV analysis. Our study expands the genetic spectrum of PCD with DNAH5 in the Chinese population.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s43657-023-00130-0.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s43657-023-00130-0.
摘要：
原发性纤毛运动障碍（PCD）是一种罕见的疾病，其特征是广泛的遗传异质性。然而,在PCD的遗传发病机制中，拷贝数变异（CNV）没有得到足够的重视，很少有报道，尤其是在中国。在高度怀疑患有PCD的患者中使用下一代测序（NGS），然后进行靶向CNV分析，常规全外显子组测序（WES）分析结果为阴性。使用定量实时聚合酶链反应(qPCR)和Sanger测序来确认这些CNV。为了进一步表征纤毛表型，高速视频显微镜分析（HSVA），透射电子显微镜（TEM），和免疫荧光（IF）分析。患者1（F1：II-1），一个0.6岁的女孩,来自一个非近亲家庭-I她提出了全situsinversustotalis，新生儿呼吸窘迫,和鼻窦炎.鼻腔一氧化氮水平明显降低。呼吸纤毛搏动幅度减小。TEM显示纤毛的外动力蛋白臂（ODA）缩短。通过基于NGS的CNV分析鉴定出跨越外显子71-72的chr5：13717907-13722661del。患者2（F2：IV-4），一个37岁的男人,和他的大哥病人3（F2：IV-2）来自一个近亲家庭-II。都有鼻窦炎,支气管扩张和全位倒置。发现患者2和患者3的呼吸纤毛均匀不运动，ODA缺陷。两个新的纯合缺失chr5：13720087_13733030delinsGTTTC和chr5：13649539_13707643del，通过基于NGS的CNV分析鉴定了跨外显子69-71和外显子77-79。通过qPCR扩增确认DNA拷贝数的异常。图IF显示患者1和患者2的呼吸纤毛缺乏动力蛋白轴突重链5(DNAH5)蛋白表达。该报告通过基于WES的CNV分析鉴定了三种新的DNAH5疾病相关变体。我们的研究扩展了具有DNAH5的PCD在中国人群中的遗传谱。
■在线版本包含补充材料，可在10.1007/s43657-023-00130-0获得。
■在线版本包含补充材料，可在10.1007/s43657-023-00130-0获得。
