Burns Randi, Majczenko Karen, Xu Jishu, Peng Weiping, Yapici Zuhal, Dowling James J, Li Jun Z, Burmeister Margit
From the Program in Cellular and Molecular Biology (R.B., M.B.), Molecular & Behavioral Neuroscience Institute (R.B., K.M., M.B.), Departments of Human Genetics (J.X., W.P., J.Z.L., M.B.), Neurology (J.J.D.), Pediatrics (J.J.D.), and Psychiatry (M.B.), University of Michigan Medical Center, Ann Arbor; and Department of Neurology (Z.Y.), Division of Child Neurology, Istanbul Faculty of Medicine, Istanbul University, Turkey. J.J.D. is currently affiliated with the Division of Neurology and Program of Genetics and Genome Biology, Hospital for Sick Children, Departments of Pediatrics and Molecular Genetics, University of Toronto, Canada.
Neurology. 2014 Dec 2;83(23):2175-82. doi: 10.1212/WNL.0000000000001053. Epub 2014 Oct 31.
To elucidate the genetic cause of a rare recessive ataxia presented by 2 siblings from a consanguineous Turkish family with a nonprogressive, congenital ataxia with mental retardation of unknown etiology.
Whole-exome sequencing was combined with homozygosity mapping, linkage, and expression analysis to identify candidate genes, confirmed by Sanger sequencing. Reverse transcription-PCR and immunoblotting were used to determine the functional consequences of the gene variant. A zebrafish model was developed using morpholino-mediated knockdown.
We identified a homozygous mutation at the invariant +1 position (c.964+1G>A) in intron 9 of the CWF19L1 (complexed with cdc5 protein 19-like 1) gene. This mutation is absent in >6,500 European and African American individuals and 200 Turkish control DNAs. The mutation causes exon skipping, reduction in messenger RNA levels, and protein loss in cell lines of affected individuals. Morpholino-mediated knockdown in a zebrafish model demonstrates that loss of the evolutionarily highly conserved CWF19L1, whose normal biological function is unknown, alters cerebellar morphology and causes movement abnormalities.
Our results suggest that CWF19L1 mutations may be a novel cause of recessive ataxia with developmental delay. Our research may help with diagnosis, especially in Turkey, identify causes of other ataxias, and may lead to novel therapies.
阐明一个来自土耳其近亲家庭的2名同胞所患罕见隐性共济失调的遗传病因,该疾病为非进行性先天性共济失调,伴有病因不明的智力发育迟缓。
全外显子组测序结合纯合性定位、连锁分析和表达分析来鉴定候选基因,并通过桑格测序进行确认。采用逆转录聚合酶链反应和免疫印迹法来确定基因变异的功能后果。利用吗啉代介导的敲低技术建立斑马鱼模型。
我们在CWF19L1(与细胞分裂周期蛋白5样蛋白1复合)基因第9内含子的不变+1位置(c.964+1G>A)发现了一个纯合突变。在6500多名欧洲和非裔美国人个体以及200份土耳其对照DNA中均未发现该突变。该突变导致外显子跳跃、信使核糖核酸水平降低以及受影响个体细胞系中的蛋白质缺失。斑马鱼模型中吗啉代介导的敲低实验表明,进化上高度保守的CWF19L1(其正常生物学功能未知)的缺失会改变小脑形态并导致运动异常。
我们的结果表明,CWF19L1突变可能是导致伴有发育迟缓的隐性共济失调的新病因。我们的研究可能有助于诊断,尤其是在土耳其,有助于确定其他共济失调的病因,并可能带来新的治疗方法。