School of Medicine, Medical Sciences and Nutrition, Institute of Medical Sciences, Foresterhill, University of Aberdeen, Aberdeen, Scotland, AB25 2ZD, UK.
Rowett Institute of Nutrition and Health, School of Medicine, Medical Sciences and Nutrition, Foresterhill, University of Aberdeen, Aberdeen, Scotland, AB25 2ZD, UK.
Mol Psychiatry. 2021 Jun;26(6):2263-2276. doi: 10.1038/s41380-020-0707-7. Epub 2020 Mar 13.
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%的死亡人数有关。然而，这一数字在男性中尤为突出，有 7.6%的死亡可归因于饮酒。先前的研究表明，在不同的男性群体中，甘丙肽（GAL）基因的基因型与焦虑和酗酒之间存在显著的相互作用，但未能确定其机制。为了解决这些问题，本研究分析了英国生物银行的人类队列，并在男性中发现了GAL5.1 增强子高度保守的人类等位基因变异（GG 或 CA 基因型）与焦虑和酒精摄入量（AUDIT 问卷评分）之间的显著相互作用（n=115865；p=0.0007）。至关重要的是，使用 CRISPR 基因组编辑技术在小鼠中破坏 GAL5.1，显著降低了杏仁核和下丘脑的 GAL 表达，同时减少了 KO 小鼠对乙醇的摄入。有趣的是，我们还发现，雄性 GAL5.1 KO 动物的焦虑样行为减少，这与我们在英国生物银行研究中观察到的人类情况相似。通过生物信息学分析和共转染研究，我们进一步确定了 EGR1 转录因子，它与杏仁核和下丘脑中的 GAL 共表达，是 GAL5.1 GG 基因型的蛋白激酶 C（PKC）支持活性的重要因素，但在 CA 基因型中则不那么重要。我们的独特研究使用了人类关联分析、CRISPR 基因组编辑在小鼠中的应用、动物行为分析和细胞培养研究的新颖组合，鉴定了一个高度保守的调节机制，将焦虑和酒精摄入量联系起来，这可能导致男性对焦虑和酗酒的易感性增加。
