<i>In Silico</i> Pharmaco-genomic Analysis and Structural Simulations Unveil Potential of Bumetanide in Mitigating NLGN3 R451C Mutation Effects in Autism Spectrum Disorder

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition influenced by genetic and epigenetic factors. This study integrates data from genome-wide association studies (GWAS), epitranscriptomic RNA modifications and machine learning techniques to analyze mutations in neuroligin-3 (NLGN3), focusing on the R451C variant. GWAS identified 90 significant single nucleotide polymorphisms (SNPs), with rs910805 being the most significant ([Formula: see text] 2.041 × 10[Formula: see text], OR [Formula: see text] 0.90874), alongside over 600 m6A-SNPs and 78 m5C-SNPs with cis-eQTL effects. Structural analysis revealed that R451C destabilizes NLGN3, increasing the root-mean-square deviation (RMSD) and root-mean-square fluctuation (RMSF), with the [Formula: see text] shifting (1.1 kcal/mol, –0.11 kcal/mol), suggesting impaired synaptic function. Molecular docking and MM (PB/GB)SA simulations identified bumetanide (–38.13 kcal/mol) as the most promising therapeutic candidate, followed by metformin (–34.25 kcal/mol) and oxytocin (–21.12 kcal/mol). This study highlights genetic, structural, neuroanatomical and pharmacological insights into ASD, supporting bumetanide as a potential therapy. Further in vivo validation is needed for clinical translation.