Next-generation sequencing identified novel Desmoplakin frame-shift variant in patients with Arrhythmogenic cardiomyopathy
Next-generation sequencing identified novel Desmoplakin frame-shift variant in patients with Arrhythmogenic cardiomyopathy
复制标题
新一代测序在致心律失常性心肌病患者中发现了新型桥粒斑蛋白移码变异体
DOI：
10.1186/s12872-020-01369-5
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发表时间：
2020-02-11
影响因子：
2.1
通讯作者：
Xiang, Meixiang
中科院分区：
文献类型：
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作者：
Lin, Xiaoping;Ma, Yuankun;Xiang, Meixiang
背景：心律失常性心肌病（AC）是心源性猝死（SCD）的主要原因之一。最近的研究发现，心脏桥粒的突变是AC发病的关键因素。然而，个体家庭的具体病因在很大程度上仍然未知。方法选取有晕厥、致死性室性心律失常和SCD的4代家族。通过Sanger测序进行靶向下一代测序（Targeted next generation sequencing， NGS）并进行验证。构建了含有突变型和野生型（WT）的质粒。采用Real-time PCR、western-blot和免疫荧光检测突变后的功能变化。结果先证者56岁，女性，表现为反复心悸和晕厥。由于她的SCD家族史/ SCD流产，植入了ICD。NGS结果显示，Desmoplakin （DSP）在5个家族成员中发现了一个新的杂合移帧变异（c.832delG）。这种变异导致帧移位和过早终止，产生一个截断的蛋白质。携带相同变体的家族成员的心脏磁共振（CMR）显示右心室心肌变薄和脂肪浸润，双心室晚期钆增强阳性，RV功能障碍严重，符合AC的诊断标准。转染突变质粒的HEK293T细胞表达截断的dspmrna和蛋白，核连接血小板红蛋白（JUP）上调，β-catenin下调。结论：我们推断小说是一种新的小说。在该家族中，832delGvariant indsp与AC相关，可能通过Wnt/β-catenin信号通路。
BackgroundArrhythmogenic cardiomyopathy (AC) is one of the leading causes for sudden cardiac death (SCD). Recent studies have identified mutations in cardiac desmosomes as key players in the pathogenesis of AC. However, the specific etiology in individual families remains largely unknown.MethodsA 4-generation family presenting with syncope, lethal ventricular arrhythmia and SCD was recruited. Targeted next generation sequencing (NGS) was performed and validated by Sanger sequencing. Plasmids containing the mutation and wild type (WT) were constructed. Real-time PCR, western-blot and immunofluorescence were performed to detect the functional change due to the mutation.ResultsThe proband, a 56-year-old female, presented with recurrent palpitations and syncope. An ICD was implanted due to her family history of SCD/ aborted SCD. NGS revealed a novel heterozygous frame-shift variant (c.832delG) in Desmoplakin (DSP) among 5 family members. The variant led to frame-shift and premature termination, producing a truncated protein. Cardiac magnetic resonance (CMR) of the family members carrying the same variant shown myocardium thinning and fatty infiltration in the right ventricular, positive bi-ventricular late gadolinium enhancement and severe RV dysfunction, fulfilling the diagnostic criteria of AC. HEK293T cells transfected with mutant plasmids expressed truncatedDSPmRNA and protein, upregulation of nuclear junction plakoglobin (JUP) and downregulation of β-catenin, when compared with WT.ConclusionWe infer that the novelc.832delGvariant inDSPwas associated with AC in this family, likely through Wnt/β-catenin signaling pathway.