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

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

doi:10.3390/ijms24119548

Open AccessCase Report

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

by

Nicolò Salvarani

^1,2,†

,

Giovanni Peretto

^3,4,†

,

Crasto Silvia

^2,‡

,

Andrea Villatore

⁴

,

Cecilia Thairi

^2,5

,

Anna Santoni

⁶,

Camilla Galli

²

,

Paola Carrera

^6,7

,

Simone Sala

^3,*

,

Sara Benedetti

⁶,

Elisa Di Pasquale

^1,2,*,§

and

Chiara Di Resta

^4,6,§

¹

Institute of Genetic and Biomedical Research (IRGB), UOS of Milan, National Research Council of Italy, 20138 Milan, Italy

²

IRCCS Humanitas Research Hospital, Via Manzoni 56, Rozzano, 20089 Milan, Italy

³

Department of Cardiac Electrophysiology and Arrhythmology, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy

⁴

Faculty of Medicine, Vita-Salute San Raffaele University, 20132 Milan, Italy

⁵

Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, Pieve Emanuele, 20072 Milan, Italy

⁶

Genomic Unit for the Diagnosis of Human Pathologies, IRCCS San Raffaele Hospital, 20132 Milan, Italy

⁷

Laboratory of Clinical Molecular Biology, IRCCS San Raffaele Hospital, 20132 Milan, Italy

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

^‡

Current address: Amarin Italy S.r.l., Corso Vercelli 40, 20145 Milan, Italy.

^§

These authors contributed equally to this work.

Int. J. Mol. Sci. 2023, 24(11), 9548; https://doi.org/10.3390/ijms24119548

Submission received: 19 April 2023 / Revised: 18 May 2023 / Accepted: 25 May 2023 / Published: 31 May 2023

(This article belongs to the Special Issue Molecular Research in Heart Disease)

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Abstract

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.

Keywords: Brugada syndrome; sodium current; cardiomyocyte

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MDPI and ACS Style

Salvarani, N.; Peretto, G.; Silvia, C.; Villatore, A.; Thairi, C.; Santoni, A.; Galli, C.; Carrera, P.; Sala, S.; Benedetti, S.; et al. Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells. Int. J. Mol. Sci. 2023, 24, 9548. https://doi.org/10.3390/ijms24119548

AMA Style

Salvarani N, Peretto G, Silvia C, Villatore A, Thairi C, Santoni A, Galli C, Carrera P, Sala S, Benedetti S, et al. Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells. International Journal of Molecular Sciences. 2023; 24(11):9548. https://doi.org/10.3390/ijms24119548

Chicago/Turabian Style

Salvarani, Nicolò, Giovanni Peretto, Crasto Silvia, Andrea Villatore, Cecilia Thairi, Anna Santoni, Camilla Galli, Paola Carrera, Simone Sala, Sara Benedetti, and et al. 2023. "Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells" International Journal of Molecular Sciences 24, no. 11: 9548. https://doi.org/10.3390/ijms24119548

APA Style

Salvarani, N., Peretto, G., Silvia, C., Villatore, A., Thairi, C., Santoni, A., Galli, C., Carrera, P., Sala, S., Benedetti, S., Di Pasquale, E., & Di Resta, C. (2023). Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells. International Journal of Molecular Sciences, 24(11), 9548. https://doi.org/10.3390/ijms24119548

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

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