Identification of a novel LMF1 nonsense mutation responsible for severe hypertriglyceridemia by targeted next-generation sequencing
Identification of a novel LMF1 nonsense mutation responsible for severe hypertriglyceridemia by targeted next-generation sequencing
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DOI：
10.1016/j.jacl.2017.01.003
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发表时间：
2017-02-01
影响因子：
4.4
通讯作者：
Averna, Maurizio R.
中科院分区：
文献类型：
--
作者：
Cefalu, Angelo B.;Spina, Rossella;Averna, Maurizio R.
背景：严重的高甘油三酯血症（HTG）可能是由影响血管内富含甘油三酯（TG）的脂蛋白脂解的基因突变引起的。目的：本研究的目的是开发以严重HTG为特征的疾病的分子诊断的靶向下一代测序面板。方法：我们为下一代测序Ion Torrent Personal Genome Machine开发了一个有针对性的定制面板，以捕获影响TG合成和代谢主要途径的18个基因的编码外显子和内含子/外显子边界。我们对11例重度HTG （TG bb0 885 mg/dL-10 mmol/L）患者样本进行了测序：4例阳性对照，其中Sanger测序已鉴定出致病突变，7例患者分子缺陷尚不清楚。结果：定制的面板是准确的，并且可以确认先前在所有原发性严重HTG阳性对照中发现的遗传变异。7例HTG患者中仅有1例被发现携带LMF1基因第三个新突变（c.1380C b> G-p.Y460X）的纯合致病突变。临床和分子家族级联筛选允许鉴定另外2个受影响的兄弟姐妹和7个突变的杂合携带者。结论：我们的研究表明，与传统的Sanger重测序相比，我们的靶向重测序方法用于重度HTG的基因诊断似乎更准确，更省时，更经济。候选基因致病性突变的鉴定仍然具有挑战性，临床重测序应主要针对具有强单基因重度HTG临床标准的患者。(C) 2017年全国血脂协会。版权所有。
BACKGROUND: Severe hypertriglyceridemia (HTG) may result from mutations in genes affecting the intravascular lipolysis of triglyceride (TG)-rich lipoproteins.OBJECTIVE: The aim of this study was to develop a targeted next-generation sequencing panel for the molecular diagnosis of disorders characterized by severe HTG.METHODS: We developed a targeted customized panel for next-generation sequencing Ion Torrent Personal Genome Machine to capture the coding exons and intron/exon boundaries of 18 genes affecting the main pathways of TG synthesis and metabolism. We sequenced 11 samples of patients with severe HTG (TG>885 mg/dL-10 mmol/L): 4 positive controls in whom pathogenic mutations had previously been identified by Sanger sequencing and 7 patients in whom the molecular defect was still unknown.RESULTS: The customized panel was accurate, and it allowed to confirm genetic variants previously identified in all positive controls with primary severe HTG. Only 1 patient of 7 with HTG was found to be carrier of a homozygous pathogenic mutation of the third novel mutation of LMF1 gene (c.1380C>G-p.Y460X). The clinical and molecular familial cascade screening allowed the identification of 2 additional affected siblings and 7 heterozygous carriers of the mutation.CONCLUSIONS: We showed that our targeted resequencing approach for genetic diagnosis of severe HTG appears to be accurate, less time consuming, and more economical compared with traditional Sanger resequencing. The identification of pathogenic mutations in candidate genes remains challenging and clinical resequencing should mainly intended for patients with strong clinical criteria for monogenic severe HTG. (C) 2017 National Lipid Association. All rights reserved.