Targeted deep sequencing reveals no definitive evidence for somatic mosaicism in atrial fibrillation.
Targeted deep sequencing reveals no definitive evidence for somatic mosaicism in atrial fibrillation.
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DOI：
10.1161/circgenetics.114.000650
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发表时间：
2015-02
期刊：
影响因子：
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通讯作者：
Marcus GM
中科院分区：
文献类型：
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作者：
Roberts JD;Longoria J;Poon A;Gollob MH;Dewland TA;Kwok PY;Olgin JE;Deo RC;Marcus GM
对≤15例房颤（AF）患者的研究发现了连接蛋白基因内的心房特异性突变，表明体细胞突变可能是偶发性心律失常病例的原因。我们试图使用560个基因的靶向深度下一代测序来识别患有和不患有AF的患者中的心房体细胞突变，包括与AF有关的遗传罪犯，孟德尔心肌病和通道病，以及基因组内的所有离子通道。对34例患者（25例AF）淋巴细胞和左心耳的DNA进行靶向基因捕获和下一代测序。20例房颤患者接受了专门用于肺静脉隔离的心脏手术，17例无结构性心脏病。使用Burrows-Wheeler Aligner、基因组分析工具包和MuTect软件包对每个心房-淋巴细胞对进行序列比对和变异识别。下一代测序产生了中位数265倍的覆盖深度（IQR 164-369）。比较来自每个心房淋巴细胞对的300万个碱基对，发现3名AF患者和2名对照中存在单个潜在体细胞错义突变（12 vs. 11%; p=1）。所有潜在的不一致变体具有低等位基因分数（范围：2.3-7.3%），并且常规测序均未检测到。使用高深度的下一代测序和最先进的体细胞突变调用方法，在一个综合性心律失常遗传小组中，25例AF患者中没有证实致病性心房体细胞突变。这些发现表明心房特异性突变是罕见的，体细胞嵌合体不太可能在AF发病机制中发挥重要作用。
Studies of ≤15 atrial fibrillation (AF) patients have identified atrial-specific mutations within connexin genes, suggesting that somatic mutations may account for sporadic cases of the arrhythmia. We sought to identify atrial somatic mutations among patients with and without AF using targeted deep next-generation sequencing of 560 genes, including genetic culprits implicated in AF, the Mendelian cardiomyopathies and channelopathies, and all ion channels within the genome. Targeted gene capture and next generation sequencing were performed on DNA from lymphocytes and left atrial appendages of 34 patients (25 with AF). Twenty AF patients had undergone cardiac surgery exclusively for pulmonary vein isolation, and 17 had no structural heart disease. Sequence alignment and variant calling were performed for each atrial-lymphocyte pair using the Burrows-Wheeler Aligner, the Genome Analysis Toolkit, and MuTect packages. Next generation sequencing yielded a median 265-fold coverage depth (IQR 164–369). Comparison of the 3 million base pairs from each atrial-lymphocyte pair revealed a single potential somatic missense mutation in 3 AF patients and 2 in a single control (12 vs. 11%; p=1). All potential discordant variants had low allelic fractions (range: 2.3–7.3%) and none were detected with conventional sequencing. Using high-depth next generation sequencing and state-of-the art somatic mutation calling approaches, no pathogenic atrial somatic mutations could be confirmed among 25 AF patients in a comprehensive cardiac arrhythmia genetic panel. These findings indicate that atrial specific mutations are rare and that somatic mosaicism is unlikely to exert a prominent role in AF pathogenesis.