CRISPR disruption and UK Biobank analysis of a highly conserved polymorphic enhancer suggests a role in male anxiety and ethanol intake

Andrew R McEwan, Connor Davidson, Elizabeth Hay, Yvonne Turnbull, Johanna Celene Erickson, Pietro Marini, Dana Wilson, Andrew McIntosh, Mark J. Adams, Chris Murgatroyd, Perry Barrett, Mirela Delibegovic, Toni-Kim Clarke, Alasdair MacKenzie* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
JournalMolecular Psychiatry
Early online date13 Mar 2020
DOIs
Publication statusPublished - 13 Mar 2020

Keywords

  • UK Biobank
  • CRISPR genome editing
  • tissue-specific enhancer
  • galanin
  • ethanol intake
  • PKC
  • EGR1
  • male anxiety
  • DEPRESSION
  • REMOTE ENHANCER
  • DISORDERS
  • DEPENDENCE
  • PARAVENTRICULAR NUCLEUS
  • GALANIN
  • GENE-EXPRESSION
  • CONSUMPTION
  • RANGE
  • HYPOTHALAMUS

ASJC Scopus subject areas

  • Psychiatry and Mental health
  • Cellular and Molecular Neuroscience
  • Molecular Biology

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