CCDC65 mutation causes primary ciliary dyskinesia with normal ultrastructure and hyperkinetic cilia.
CCDC65 mutation causes primary ciliary dyskinesia with normal ultrastructure and hyperkinetic cilia.
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DOI：
10.1371/journal.pone.0072299
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kerem E
中科院分区：
文献类型：
--
作者：
Horani A;Brody SL;Ferkol TW;Shoseyov D;Wasserman MG;Ta-shma A;Wilson KS;Bayly PV;Amirav I;Cohen-Cymberknoh M;Dutcher SK;Elpeleg O;Kerem E
原发性纤毛运动障碍（PCD）是一种遗传性疾病，其特征是纤毛功能受损，导致慢性鼻窦炎。PCD的遗传原因仍在不断发展，而诊断通常依赖于发现纤毛超微结构异常和不动纤毛。在这里，我们报告了一个新的基因与PCD，但没有纤毛超微结构异常明显的透射电子显微镜，但与运动障碍纤毛跳动。在一个PCD受试者的家庭中进行遗传连锁分析。基因表达研究在衣原体和人类气道上皮细胞，使用RNA检测和免疫染色。候选基因突变的表型效应在用基因靶向的shRNA序列转导的原代培养人气管支气管上皮细胞中确定。视频显微镜用于评估纤毛运动。一个单一的新突变CCDC 65，它创建了一个终止密码子在位置293，被确定在一个受试者与典型的PCD临床特征。CCDC 65是衣原体连接蛋白-动力蛋白调节复合物蛋白DRC 2的直向同源物，定位于正常鼻上皮细胞的纤毛，但在先证者中不存在。在培养的气道上皮细胞中，CCDC 65的表达在纤毛发生过程中上调，在C.莱因哈德氏菌与对照细胞相比，受影响个体的鼻上皮细胞和CCDC 65特异性shRNA转导的正常气道上皮细胞具有僵硬和运动障碍的纤毛跳动模式。此外，Gas 8，一种先前鉴定为与CCDC 65相关的连接蛋白-动力蛋白调节复合物组分，在PCD受试者和CCDC 65沉默细胞的气道细胞中不存在。连接蛋白-动力蛋白调控复合体成员CCDC 65的突变导致移码突变和PCD。受影响的个人有改变纤毛跳动模式，并没有检测到的纤毛轴丝超微结构缺陷，强调的作用，连接蛋白-动力蛋白调节复合物和某些方法的局限性PCD诊断。
Primary ciliary dyskinesia (PCD) is a genetic disorder characterized by impaired ciliary function, leading to chronic sinopulmonary disease. The genetic causes of PCD are still evolving, while the diagnosis is often dependent on finding a ciliary ultrastructural abnormality and immotile cilia. Here we report a novel gene associated with PCD but without ciliary ultrastructural abnormalities evident by transmission electron microscopy, but with dyskinetic cilia beating. Genetic linkage analysis was performed in a family with a PCD subject. Gene expression was studied in Chlamydomonas reinhardtii and human airway epithelial cells, using RNA assays and immunostaining. The phenotypic effects of candidate gene mutations were determined in primary culture human tracheobronchial epithelial cells transduced with gene targeted shRNA sequences. Video-microscopy was used to evaluate cilia motion. A single novel mutation in CCDC65, which created a termination codon at position 293, was identified in a subject with typical clinical features of PCD. CCDC65, an orthologue of the Chlamydomonas nexin-dynein regulatory complex protein DRC2, was localized to the cilia of normal nasal epithelial cells but was absent in those from the proband. CCDC65 expression was up-regulated during ciliogenesis in cultured airway epithelial cells, as was DRC2 in C. reinhardtii following deflagellation. Nasal epithelial cells from the affected individual and CCDC65-specific shRNA transduced normal airway epithelial cells had stiff and dyskinetic cilia beating patterns compared to control cells. Moreover, Gas8, a nexin-dynein regulatory complex component previously identified to associate with CCDC65, was absent in airway cells from the PCD subject and CCDC65-silenced cells. Mutation in CCDC65, a nexin-dynein regulatory complex member, resulted in a frameshift mutation and PCD. The affected individual had altered cilia beating patterns, and no detectable ultrastructural defects of the ciliary axoneme, emphasizing the role of the nexin-dynein regulatory complex and the limitations of certain methods for PCD diagnosis.
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影响因子：
4.8
作者：
Lin, Jianfeng;Tritschler, Douglas;Nicastro, Daniela
通讯作者：
Nicastro, Daniela
影响因子：
9.8
作者：
Knowles, Michael R.;Leigh, Margaret W.;Zariwala, Maimoona A.
通讯作者：
Zariwala, Maimoona A.
影响因子：
9.8
作者：
Edvardson, Simon;Shaag, Avraham;Elpeleg, Orly
通讯作者：
Elpeleg, Orly
影响因子：
9.8
作者：
Guichard, C;Harricane, MC;Bouvagnet, P
通讯作者：
Bouvagnet, P
影响因子：
--
作者：
Bekker, Janine M.;Colantonio, Jessica R.;Crosbie, Rachelle H.
通讯作者：
Crosbie, Rachelle H.