Department of Human Genetics, Emory University School of Medicine, Atlanta, GA 30322, USA.
Am J Hum Genet. 2012 Feb 10;90(2):363-8. doi: 10.1016/j.ajhg.2011.12.024. Epub 2012 Feb 2.
Congenital disorders of glycosylation (CDG) are inherited autosomal-recessive diseases that impair N-glycosylation. Approximately 20% of patients do not survive beyond the age of 5 years old as a result of widespread organ dysfunction. Although most patients receive a CDG diagnosis based on abnormal glycosylation of transferrin, this test cannot provide a genetic diagnosis; indeed, many patients with abnormal transferrin do not have mutations in any known CDG genes. Here, we combined biochemical analysis with whole-exome sequencing (WES) to identify the genetic defect in an untyped CDG patient, and we found a 22 bp deletion and a missense mutation in DDOST, whose product is a component of the oligosaccharyltransferase complex that transfers the glycan chain from a lipid carrier to nascent proteins in the endoplasmic reticulum lumen. Biochemical analysis with three biomarkers revealed that N-glycosylation was decreased in the patient's fibroblasts. Complementation with wild-type-DDOST cDNA in patient fibroblasts restored glycosylation, indicating that the mutations were pathological. Our results highlight the power of combining WES and biochemical studies, including a glyco-complementation system, for identifying and confirming the defective gene in an untyped CDG patient. This approach will be very useful for uncovering other types of CDG as well.
先天性糖基化障碍(CDG)是一种遗传性常染色体隐性疾病,会损害 N-糖基化。由于广泛的器官功能障碍,约 20%的患者在 5 岁之前无法存活。尽管大多数患者根据转铁蛋白的异常糖基化被诊断为 CDG,但该检测无法提供基因诊断;实际上,许多转铁蛋白异常的患者没有任何已知 CDG 基因的突变。在这里,我们将生化分析与外显子组测序(WES)相结合,以确定未分型 CDG 患者的遗传缺陷,并在 DDOST 中发现了 22 个碱基对的缺失和错义突变,其产物是寡糖基转移酶复合物的一部分,该复合物将聚糖链从脂质载体转移到内质网腔中新生蛋白质上。使用三种生物标志物的生化分析表明,患者成纤维细胞中的 N-糖基化减少。在患者成纤维细胞中用野生型 DDOST cDNA 进行互补恢复了糖基化,表明突变是病理性的。我们的结果强调了将 WES 与生化研究(包括糖基化互补系统)相结合,用于识别和确认未分型 CDG 患者缺陷基因的强大功能。这种方法对于揭示其他类型的 CDG 也将非常有用。