Despite a significant genetic component to insomnia (heritability: 22%-25%), the genetic loci that modulate insomnia risk remain limited. We used the Unified Test for Molecular Markers (UTMOST) for transcriptome-wide association studies (TWAS) across various tissues, integrating summary statistics from a Genome-Wide Association Study (GWAS) of 462,341 European participants with gene expression data from the Genotype-Tissue Expression (GTEx) project. Three validation methods (FUSION, FOCUS, and MAGMA) were used to confirm important genes. Tissue and functional enrichment analyses of insomnia-related single-nucleotide polymorphisms (SNPs) were conducted with MAGMA. Conditional and joint analyses, along with fine mapping, were used to enhance our understanding of insomnia's genetic architecture. Mendelian randomization was used to assess causal associations between significant genes and insomnia. Two novel susceptibility genes,and, were identified as linked to insomnia risk using four TWAS approaches. Mendelian randomization analysis suggestsincreases the risk of insomnia. Tissue enrichment analyses indicated that insomnia-related SNPs were enriched in specific brain regions, including the cerebellum, frontal cortex (BA9), hypothalamus, and hippocampus. Conditional and joint analyses identified two genomic regions (2p16.1 and 4q22.1). Functional enrichment analyses showed that pathways related to insomnia involve the SMAD2/3 pathway, synaptic function, and oxidative stress. This study identifies two new candidate genes,and, that may contribute to insomnia risk through neurodevelopment, neuroinflammation, and synaptic function, suggesting potential therapeutic targets.This study identifiesandas novel susceptibility genes for insomnia through transcriptome-wide association studies (TWAS). Mendelian randomization confirms a causal link betweenand insomnia. Key brain regions, including the cerebellum and frontal cortex, and critical pathways like SMAD2/3 signaling and oxidative stress are implicated. These findings provide new insights into the genetic basis of insomnia. VRK2MMRN1VRK2VRK2MMRN1VRK2MMRN1VRK2NEW & NOTEWORTHY
