Brugada综合征(BrS)是一种遗传的常染色体显性遗传心脏通道病。SCN5A基因的致病性罕见突变,编码电压依赖性心脏Na+通道蛋白(Nav1.5)的α亚基,在20%的BrS患者中被发现,影响通道的正确功能。迄今为止,尽管数百种SCN5A变种与BrS相关,在大多数情况下,潜在的致病机制仍不清楚。因此,SCN5ABrS罕见变异体的功能特征仍然是一个主要障碍,是确认其致病作用的基础.由多能干细胞(PSC)分化的人心肌细胞(CM)已被广泛证明是研究心脏病的可靠平台。能够概括疾病的特定特征,包括心律失常事件和传导异常。基于此,在这项研究中,我们对BrS家族性罕见变异NM_198056.2:c.3673G>A(NP_932173.1:p。Glu1225Lys),以前从未在心脏相关的背景下进行功能表征,作为人类心肌细胞。使用编码GFP标记的SCN5A基因的特定慢病毒载体,该基因携带特定的c.3673G>A变体和与对照PSC(PSC-CM)分化的CM,我们证明了突变的Nav1.5受损,因此表明罕见的BrS检测变体的致病性。更广泛地说,我们的工作支持PSC-CMs用于评估基因变异的致病性,由于下一代测序方法的进步及其在基因检测中的大量使用,其鉴定呈指数级增长。
Brugada syndrome (BrS) is an inherited autosomal dominant cardiac channelopathy. Pathogenic rare mutations in the SCN5A gene, encoding the alpha-subunit of the voltage-dependent cardiac Na+ channel protein (Nav1.5), are identified in 20% of BrS patients, affecting the correct function of the channel. To date, even though hundreds of SCN5A variants have been associated with BrS, the underlying pathogenic mechanisms are still unclear in most cases. Therefore, the functional characterization of the SCN5A BrS rare variants still represents a major hurdle and is fundamental to confirming their pathogenic effect. Human cardiomyocytes (CMs) differentiated from pluripotent stem cells (PSCs) have been extensively demonstrated to be reliable platforms for investigating cardiac diseases, being able to recapitulate specific traits of disease, including arrhythmic events and conduction abnormalities. Based on this, in this study, we performed a functional analysis of the BrS familial rare variant NM_198056.2:c.3673G>A (NP_932173.1:p.Glu1225Lys), which has been never functionally characterized before in a cardiac-relevant context, as the human
cardiomyocyte. Using a specific lentiviral vector encoding a GFP-tagged SCN5A gene carrying the specific c.3673G>A variant and CMs differentiated from control PSCs (PSC-CMs), we demonstrated an impairment of the mutated Nav1.5, thus suggesting the pathogenicity of the rare BrS detected variant. More broadly, our work supports the application of PSC-CMs for the assessment of the pathogenicity of gene variants, the identification of which is increasing exponentially due to the advances in next-generation sequencing methods and their massive use in genetic testing.