Abnormal glycosylation in Joubert syndrome type 10.
Abnormal glycosylation in Joubert syndrome type 10.
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DOI：
10.1186/s13630-017-0048-6
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Boerkoel, Cornelius F
中科院分区：
文献类型：
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作者：
Kane, Megan S;Davids, Mariska;Boerkoel, Cornelius F
背景技术背景：疾病发病机制的发现需要系统的不可知筛选的多个稳态过程，可能成为失调。我们说明了这一原则的评估和诊断的一个5岁的男孩与Joubert综合征10型（JBTS 10）。他携带OFD 1突变p.Gln886Lysfs*2（NM_003611.2：c.2656del），表现出Joubert综合征的特征。方法：我们整合了外显子组测序，血浆和培养的皮肤成纤维细胞糖组的MALDI-TOF质谱分析，以及对先证者的全面临床评估。通过免疫荧光法进行纤毛形成和凝集素染色的分析。细胞核苷酸糖水平的测量进行了高效阴离子交换色谱脉冲安培检测。统计分析利用学生和Fisher的精确t检验。结果：糖组分析血浆和培养的皮肤成纤维细胞确定异常的N-和O-连接的糖基化配置文件。这些发现在两个不相关的OFD 1突变的男性中重复。从受影响的个人培养成纤维细胞有缺陷的纤毛。先证者的成纤维细胞还具有异常升高的核唾液酸化特征和增加的总细胞CMP-唾液酸水平。结论：野生型OFD 1基因的表达对纤毛发生和糖基化异常的挽救作用提示两者均参与了JBTS 10的发病机制。
BACKGROUND: The discovery of disease pathogenesis requires systematic agnostic screening of multiple homeostatic processes that may become deregulated. We illustrate this principle in the evaluation and diagnosis of a 5-year-old boy with Joubert syndrome type 10 (JBTS10). He carried the OFD1 mutation p.Gln886Lysfs*2 (NM_003611.2: c.2656del) and manifested features of Joubert syndrome.METHODS: We integrated exome sequencing, MALDI-TOF mass spectrometry analyses of plasma and cultured dermal fibroblasts glycomes, and full clinical evaluation of the proband. Analyses of cilia formation and lectin staining were performed by immunofluorescence. Measurement of cellular nucleotide sugar levels was performed with high-performance anion-exchange chromatography with pulsed amperometric detection. Statistical analyses utilized the Student's and Fisher's exact t tests.RESULTS: Glycome analyses of plasma and cultured dermal fibroblasts identified abnormal N- and O-linked glycosylation profiles. These findings replicated in two unrelated males with OFD1 mutations. Cultured fibroblasts from affected individuals had a defect in ciliogenesis. The proband's fibroblasts also had an abnormally elevated nuclear sialylation signature and increased total cellular levels of CMP-sialic acid. Ciliogenesis and each glycosylation anomaly were rescued by expression of wild-type OFD1.CONCLUSIONS: The rescue of ciliogenesis and glycosylation upon reintroduction of WT OFD1 suggests that both contribute to the pathogenesis of JBTS10.