Characterization of non-coding genetic vairants associated with Brugada Syndrome: research and communication

Brugada syndrome (BrS) is defined as a cardiac disorder associated with high risk of sudden cardiac death and ventricular fibrillation. It is responsible of 4% of all sudden deaths and 20% of sudden cardiac deaths in patients without structural abnormalities. BrS is caused by dysfunction of ion channels involved in generation of the cardiac action potential. Approximately 20-25% of BrS patients have mutations in the coding region of the SCN5A gene, which encodes the α-subunit of the cardiac voltage-gated sodium channel. This work is included in the “Regulome-Seq” project, which aims to study non-coding DNA variants within transcription factor (TF) binding sites nearby BrS-associated genes. To characterize genetic variation in these regions, a set of pre-selected 1,293 loci was sequenced in a cohort of 89 diagnosed-BrS cases. To evaluate the consequences of the variants identified that overlap TF CTCF binding sites, the DeepBind algorithm was used to predict the effect of each variant in CTCF binding. 34 of these variants overlapping CTCF binding sites were cloned into a luciferase reporter vector to evaluate their effect in cell-based assays. After co-transfection in H9c2 cells, the luciferase activity obtained was measured to examine the CTCF binding level for each sequence. The results obtained indicate that DeepBind algorithm and luciferase assays results have a similar trend to evaluate the effect of the variants on CTCF binding. However, the results did not show a significant correlation between both methods. This suggests that the use of other DeepBind algorithms based on Machine Learning to predict BrS effects should be further explored. The experimental part of this work was complemented by its communication to the public and by innovative steps in general Science dissemination, with the conception of a Molecular Biology workshop on possible mutations in DNA. Communicating genome and DNA-related concepts is a great opportunity to improve scientific culture of society and to increase Science awareness among young students. Most people have a small idea about what BrS is, because they think is “something” related to mutations in their body hearts. This study seeks to explain the effect and influence of mutations in genetic diseases, to contribute to the open scientific attitude in the improvement of the educational process of Science in secondary education students and societ