原发性纤毛运动障碍(PCD)是一种罕见的遗传性疾病,其特征是活动纤毛功能障碍和超微结构受损。尽管有大量的研究,约30%的PCD病例的遗传基础仍有待阐明。这里,我们提出了编码卷曲螺旋结构域含蛋白40(CCDC40)的基因中的两个新突变的鉴定和功能分析,在家族性PCD病例中发现。这些新的CCDC40突变,NM_017950.4:c.2236-2delA和c.2042_2046delTCACA,NP_060420.2:p.(Ile681fs),通过全外显子组测序(WES)鉴定。然后进行Sanger测序以确认WES结果并确定先证者亲本的CCDC40基因序列。c.2042_2046delTCACA突变破坏了蛋白质的阅读框,因此预测会产生非功能性蛋白质。使用pcDNA3.1()质粒的小基因测定法,我们进一步研究了c.2236-2delA突变的潜在致病作用,发现该突变通过剪接破坏导致截短蛋白的形成.因此,总之,我们确定了CCDC40基因的两个突变,在家族性PCD病例中可以认为是致病性复合杂合突变,从而扩大了该疾病中CCDC40基因的已知突变谱。
Primary ciliary dyskinesia (PCD) is a rare genetic disorder characterized by motile ciliary dysfunction and impaired ultrastructure. Despite numerous studies, the genetic basis for about 30% of PCD cases remains to be elucidated. Here, we present the identification and functional analysis of two novel mutations in the gene encoding coiled-coil domain-containing protein 40 (CCDC40), which are found in a familial
case of PCD. These novel CCDC40 mutations, NM_017950.4: c.2236-2delA and c.2042_2046delTCACA, NP_060420.2: p.(Ile681fs), were identified by whole-exome sequencing (WES). Sanger sequencing was then performed to confirm the WES results and determine the CCDC40 gene sequences of the proband\'s parents. The c.2042_2046delTCACA mutation disrupts the reading frame of the protein and is therefore predicted to produce a non-functional protein. Using a minigene assay with the pcDNA3.1(+) plasmid, we further investigated the potential pathogenic effects of the c.2236-2delA mutation and found that this mutation leads to formation of a truncated protein via splicing disruption. Thus, in summary, we identified two mutations of the CCDC40 gene that can be considered pathogenic compound heterozygous mutations in a
case of familial PCD, thereby expanding the known mutational spectrum of the CCDC40 gene in this disease.