THE CONNECTION BETWEEN AUTISM SPECTRUM DISORDER AND THE SCN2A AND RENL GENES

Abstract

<h3>Background</h3> The incidence of autistic spectrum disorder (ASD) is growing, but the pathophysiology and the etiology is still uncertain. This disorder is caused by genetic and environmental factors causality, with the SCN2A and RELN genes being the most prominent associated to ASD. Hence, this study aims to investigate the role of SCN2A and RELN genes at ASD phenotype expression. <h3>Methods</h3> Previously produced induced pluripotent stem cells (iPS) lineages from healthy donors (EA1, EB4), SCN2A gene knockout (EB4CRISPR) and autistic patient (iM5) with this mutation proceed to cerebral organoids and neurospheres generation. Following maturation, their immunofluorescence analysis were performed. For statistical assay were used One-way ANOVA test with Tukey‘s post-test for multiple comparisons. <h3>Results</h3> iM5 embryoid bodies didn't develop cerebral organoids, in contrast to EB4 and EA1. All NSCs were cell-type validated with Q-PCR, indicating higher iM5 differentiation. iM5 neurospheres had atypical morphology and smaller neuronal extensions when compared with others at immunofluorescence. Also, knockout clone kept migrating and growing more than iM5 one – this clone also had a mutation at genes like RELN. At last, iM5 cultivated with conditioned medium (50% neurospheres basal medium and 50% medium collected from other neurospheres) from knockout clone showed better growing and migration, compared with healthy cells conditioned or basal mediums iM5 cultures. <h3>Conclusion</h3> Normal migration of knockout clone when compared with impaired one from iM5, improoved with conditionated medium, indicates that SCN2A doesn't have a main role at neuronal migration and leads to hypothesys that RELN is related to neuronal migration and growth. More experiments are needed to confirm this results.