Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells

• Published in: International journal of molecular sciences Vol. 24; no. 11; p. 9548
• Main Authors: Salvarani, Nicolò, Peretto, Giovanni, Silvia, Crasto, Villatore, Andrea, Thairi, Cecilia, Santoni, Anna, Galli, Camilla, Carrera, Paola, Sala, Simone, Benedetti, Sara, Di Pasquale, Elisa, Di Resta, Chiara
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 31-05-2023; MDPI
• Subjects: Abnormalities; Academic disciplines; Automation; Brugada syndrome; Cardiac arrhythmia; cardiomyocyte; Cardiomyocytes; Case Report; Cell differentiation; Channelopathy; Coronary artery disease; Disease; Functional analysis; Genes; Genetic screening; Genetic testing; Heart diseases; Kinases; Mutagenesis; Mutation; Next-generation sequencing; Pathogenicity; Patients; Pluripotency; Proteins; Sodium channels (voltage-gated); sodium current; Stem cells; United States > US; Italy; Brugada syndrome; sodium current; cardiomyocyte
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24119548

Brugada syndrome (BrS) is an inherited autosomal dominant cardiac channelopathy. Pathogenic rare mutations in the
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
variants have been associated with BrS, the underlying pathogenic mechanisms are still unclear in most cases. Therefore, the functional characterization of the
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
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.