In addition, the present study provided evidence that brain miRNA–mRNA regulatory networks consisting of dysregulated and negatively correlated miRNA–mRNA pairs contribute to the risk of AUD. We identified at least 17 canonical pathways that were likely influenced by dysregulated and negatively correlated miRNA–mRNA pairs in the brains of AUD subjects (Table S8). The top three pathways potentially regulated by dysregulated and negatively correlated miRNA–mRNA pairs in multiple brain regions of AUD subjects included CREB Signaling in Neurons, IL-8 Signaling, and Axonal Guidance Signaling. The CREB Signaling was found to be a central amygdaloid signaling pathway involved in high anxiety-like and excessive alcohol drinking behaviors [46]. We found that the CREB Signaling pathway could be regulated by dysregulated and negatively correlated miRNA–miRNA pairs in seven of the eight brain regions (except PUT) of AUD subjects (Figs. 4 and 5). Regarding the 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
