relationship of the IL-8 Signaling pathway and AUD, there is emerging evidence that alcohol use can stimulate immune cells to secrete peripheral pro- and anti-inflammatory cytokines (such as IL-8) [47, 48], thus supporting the role of the immune system in the pathophysiology of AUD. We observed that the IL-8 Signaling pathway could be regulated by dysregulated and negatively correlated miRNA–miRNA pairs in four (AMY, PFC, PUT, and VTA) of the eight brain regions of AUD subjects (Figs. 4 and 5). The Axon Guidance Signaling pathway can regulate axon guidance, synaptogenesis, and cell migration. Studies have shown that ethanol disrupted axon outgrowth by influencing the Axon Guidance Signaling pathway [49]. We noticed that the Axon Guidance Signaling pathway could be regulated by dysregulated and negatively correlated miRNA–miRNA pairs in four (AMY, CN, HIP, and PFC) of the eight brain regions of AUD subjects (Figs. 4 and 5). These three top pathways were validated in the Set 2 brain tissue sample by the network analysis of dysregulated and negatively correlated miRNA–mRNA pairs in six of the eight brain regions of AUD subjects (Figs. S13 and S14). Although these three top pathways were not found to be regulated by differentially expressed and negatively
