Next-generation Sequencing of RYR1 and CACNA1S in Malignant Hyperthermia and Exertional Heat Illness.
Next-generation Sequencing of RYR1 and CACNA1S in Malignant Hyperthermia and Exertional Heat Illness.
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DOI：
10.1097/aln.0000000000000610
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发表时间：
2015-05
期刊：
影响因子：
8.8
通讯作者：
Hopkins PM
中科院分区：
文献类型：
--
作者：
Fiszer D;Shaw MA;Fisher NA;Carr IM;Gupta PK;Watkins EJ;Roiz de Sa D;Kim JH;Hopkins PM
RYR 1的变异体与大多数恶性高热（MH）病例相关，这是一种由全身麻醉剂药物遗传学触发的热病形式，它们也与劳力性热病相关。CACNA 1 S也与MH有关。我们应用靶向的下一代测序方法来鉴定诊断为MH易感性的无关患者队列中RYR 1和CACNA 1 S的变体。我们还提供了第一个全面的报告测序这两个基因在一个队列的幸存者劳累性热病。使用“长”聚合酶链反应技术，在Illumina GAII®或MiSeq®平台（Illumina Inc.，加利福尼亚州圣地亚哥）。使用生物信息学方法评估变体的致病性。为了进一步随访，检测了来自其他家庭成员和多达211个MH正常和556个MH易感无关个体的DNA。在29例MH患者中，我们确定了三种致病性和四种新的RYR 1变体，另外五种RYR 1变体先前报道与MH相关。沿着在劳累性中暑队列（n = 28）中发现了三种新的RYR 1变异体，以及先前报道的两种与MH相关的变异体。另外两种变异先前被报道与中枢性肌病相关。我们分别在MH和劳力性中暑队列中发现了CACNA 1 S中一种和三种意义不明的罕见变异。靶向的下一代测序被证明有效地识别诊断有用的和潜在的牵连变异RYR 1和CACNA 1 S在MH和劳累性热病。
Variants in RYR1 are associated with the majority of cases of malignant hyperthermia (MH), a form of heat illness pharmacogenetically triggered by general anesthetics, and they have also been associated with exertional heat illness. CACNA1S has also been implicated in MH. We applied a targeted next generation sequencing approach to identify variants in RYR1 and CACNA1S in a cohort of unrelated patients diagnosed with MH susceptibility. We also provide the first comprehensive report of sequencing of these two genes in a cohort of survivors of exertional heat illness. DNA extracted from blood was genotyped using a “long” polymerase chain reaction technique, with sequencing on the Illumina GAII® or MiSeq® platforms (Illumina Inc., San Diego, CA). Variants were assessed for pathogenicity using bioinformatic approaches. For further follow up DNA from additional family members and up to 211 MH normal and 556 MH susceptible unrelated individuals was tested. In 29 MH patients we identified three pathogenic and four novel RYR1 variants, with a further five RYR1 variants previously reported in association with MH. Three novel RYR1 variants were found in the exertional heat illness cohort (n = 28) along with two more previously reported in association with MH. Two other variants were reported previously associated with centronuclear myopathy. We found one and three rare variants of unknown significance in CACNA1S in the MH and exertional heat illness cohorts respectively. Targeted next generation sequencing proved efficient at identifying diagnostically useful and potentially implicated variants in RYR1 and CACNA1S in MH and exertional heat illness.