Excessive alcohol intake is associated with 5.9% of global deaths. However, this figure is especially acute in men such that 7.6% of deaths can be attributed to alcohol intake. Previous studies identified a significant interaction between genotypes of the galanin (GAL) gene with anxiety and alcohol abuse in different male populations but were unable to define a mechanism. To address these issues the current study analysed the human UK Biobank cohort and identified a significant interaction (n = 115,865; p = 0.0007) between allelic variation (GG or CA genotypes) in the highly conserved human GAL5.1 enhancer, alcohol intake (AUDIT questionnaire scores) and anxiety in men. Critically, disruption of GAL5.1 in mice using CRISPR genome editing significantly reduced GAL expression in the amygdala and hypothalamus whilst producing a corresponding reduction in ethanol intake in KO mice. Intriguingly, we also found the evidence of reduced anxiety-like behaviour in male GAL5.1KO animals mirroring that seen in humans from our UK Biobank studies. Using bioinformatic analysis and co-transfection studies we further identified the EGR1 transcription factor, that is co-expressed with GAL in amygdala and hypothalamus, as being important in the protein kinase C (PKC) supported activity of the GG genotype of GAL5.1 but less so in the CA genotype. Our unique study uses a novel combination of human association analysis, CRISPR genome editing in mice, animal behavioural analysis and cell culture studies to identify a highly conserved regulatory mechanism linking anxiety and alcohol intake that might contribute to increased susceptibility to anxiety and alcohol abuse in men.

过量饮酒与全球 5.9% 的死亡有关。然而，这个数字在男性中尤为严重，因此 7.6% 的死亡可归因于酒精摄入。先前的研究确定了不同男性人群中甘丙肽 ( GAL)基因的基因型与焦虑和酗酒之间的显着相互作用，但无法确定机制。为了解决这些问题，目前的研究分析了人类英国生物库队列并确定了显着的相互作用（n  = 115,865；p = 0.0007) 高度保守的人类 GAL5.1 增强子中的等位基因变异（GG 或 CA 基因型）、酒精摄入量（AUDIT 问卷评分）和男性焦虑之间。至关重要的是，使用 CRISPR 基因组编辑破坏小鼠中的 GAL5.1 显着降低了杏仁核和下丘脑中GAL的表达，同时相应地减少了 KO 小鼠的乙醇摄入量。有趣的是，我们还发现了雄性 GAL5.1KO 动物焦虑样行为减少的证据，这与我们在英国生物库研究中看到的人类相似。使用生物信息学分析和共转染研究，我们进一步鉴定了与GAL共表达的 EGR1 转录因子在杏仁核和下丘脑中，在蛋白激酶 C (PKC) 中很重要，支持 GAL5.1 的 GG 基因型的活性，但在 CA 基因型中不太重要。我们独特的研究使用人类关联分析、小鼠 CRISPR 基因组编辑、动物行为分析和细胞培养研究的新组合来确定一种高度保守的调节机制，该机制将焦虑和酒精摄入联系起来，这可能有助于增加男性对焦虑和酒精滥用的易感性.