Disruption of an enhancer associated with addictive behaviour within the cannabinoid receptor-1 gene suggests a possible role in alcohol intake, cannabinoid response and anxiety-related behaviour

Elizabeth A. Hay, Andrew McEwan, Dana Wilson, Perry Barrett, Giuseppe D'Agostino, Roger G. Pertwee, Alasdair MacKenzie* (Corresponding Author)

*Corresponding author for this work

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Abstract

The cannabinoid-1 receptor (CB1) plays a critical role in a number of biological processes including nutrient intake, addiction and anxiety-related behaviour. Numerous studies have shown that expression of the gene encoding CB1 (CNR1) is highly dynamic with changes in the tissue specific expression of CNR1 associated with brain homeostasis and disease progression. However, little is known of the mechanisms regulating this dynamic expression. To gain a better understanding of the genomic mechanisms modulating the expression of CNR1 in health and disease we characterised the role of a highly conserved regulatory sequence (ECR1) in CNR1 intron 2 that contained a polymorphism in linkage disequilibrium with disease associated SNPs. We used CRISPR/CAS9 technology to disrupt ECR1 within the mouse genome. Disruption of ECR1 significantly reduced CNR1 expression in the hippocampus but not in the hypothalamus. These mice also displayed an altered sex-specific anxiety-related behavioural profile (open field test), reduced ethanol intake and a reduced hypothermic response following CB1 agonism. However, no significant changes in feeding patterns were detected. These data suggest that, whilst not all of the expression of CNR1 is modulated by ECR1, this highly conserved enhancer is required for appropriate physiological responses to a number of stimuli. The combination of comparative genomics and CRISPR/CAS9 disruption used in our study to determine the functional effects of genetic and epigenetic changes on the activity of tissue-specific regulatory elements at the CNR1 locus represent an important first step in gaining a mechanistic understanding of cannabinoid regulatory pharmacogenetics.

Original languageEnglish
Article number104407
Pages (from-to)104407
Number of pages8
JournalPsychoneuroendocrinology
Volume109
Early online date13 Aug 2019
DOIs
Publication statusPublished - Nov 2019

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Addictive Behavior
Cannabinoid Receptors
Cannabinoids
Clustered Regularly Interspaced Short Palindromic Repeats
Anxiety
Alcohols
Genes
Biological Phenomena
Conserved Sequence
Pharmacogenetics
Linkage Disequilibrium
Brain Diseases
Feeding Behavior
Genomics
Epigenomics
Introns
Hypothalamus
Single Nucleotide Polymorphism
Disease Progression
Hippocampus

Keywords

  • Cannabinoid-1 receptor
  • CRISPR genome editing
  • gene regulation
  • tissue specific
  • enhancer
  • polymorphisms
  • ethanol intake
  • CB1 agonists
  • Win55
  • 212-2
  • pharmacogenetics
  • Ethanol intake
  • Pharmacogenetics
  • Enhancer
  • NICOTINE
  • Gene regulation
  • Tissue specific
  • ENDOCANNABINOID SYSTEM
  • Anxiety-related behavior
  • Polymorphisms
  • HIPPOCAMPUS
  • Wln55,212-2
  • CB1 RECEPTORS
  • HYPOTHALAMUS

ASJC Scopus subject areas

  • Endocrine and Autonomic Systems
  • Psychiatry and Mental health
  • Biological Psychiatry
  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

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title = "Disruption of an enhancer associated with addictive behaviour within the cannabinoid receptor-1 gene suggests a possible role in alcohol intake, cannabinoid response and anxiety-related behaviour",
abstract = "The cannabinoid-1 receptor (CB1) plays a critical role in a number of biological processes including nutrient intake, addiction and anxiety-related behaviour. Numerous studies have shown that expression of the gene encoding CB1 (CNR1) is highly dynamic with changes in the tissue specific expression of CNR1 associated with brain homeostasis and disease progression. However, little is known of the mechanisms regulating this dynamic expression. To gain a better understanding of the genomic mechanisms modulating the expression of CNR1 in health and disease we characterised the role of a highly conserved regulatory sequence (ECR1) in CNR1 intron 2 that contained a polymorphism in linkage disequilibrium with disease associated SNPs. We used CRISPR/CAS9 technology to disrupt ECR1 within the mouse genome. Disruption of ECR1 significantly reduced CNR1 expression in the hippocampus but not in the hypothalamus. These mice also displayed an altered sex-specific anxiety-related behavioural profile (open field test), reduced ethanol intake and a reduced hypothermic response following CB1 agonism. However, no significant changes in feeding patterns were detected. These data suggest that, whilst not all of the expression of CNR1 is modulated by ECR1, this highly conserved enhancer is required for appropriate physiological responses to a number of stimuli. The combination of comparative genomics and CRISPR/CAS9 disruption used in our study to determine the functional effects of genetic and epigenetic changes on the activity of tissue-specific regulatory elements at the CNR1 locus represent an important first step in gaining a mechanistic understanding of cannabinoid regulatory pharmacogenetics.",
keywords = "Cannabinoid-1 receptor, CRISPR genome editing, gene regulation, tissue specific, enhancer, polymorphisms, ethanol intake, CB1 agonists, Win55, 212-2, pharmacogenetics, Ethanol intake, Pharmacogenetics, Enhancer, NICOTINE, Gene regulation, Tissue specific, ENDOCANNABINOID SYSTEM, Anxiety-related behavior, Polymorphisms, HIPPOCAMPUS, Wln55,212-2, CB1 RECEPTORS, HYPOTHALAMUS",
author = "Hay, {Elizabeth A.} and Andrew McEwan and Dana Wilson and Perry Barrett and Giuseppe D'Agostino and Pertwee, {Roger G.} and Alasdair MacKenzie",
note = "Copyright {\circledC} 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.",
year = "2019",
month = "11",
doi = "10.1016/j.psyneuen.2019.104407",
language = "English",
volume = "109",
pages = "104407",
journal = "Psychoneuroendocrinology",
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T1 - Disruption of an enhancer associated with addictive behaviour within the cannabinoid receptor-1 gene suggests a possible role in alcohol intake, cannabinoid response and anxiety-related behaviour

AU - Hay, Elizabeth A.

AU - McEwan, Andrew

AU - Wilson, Dana

AU - Barrett, Perry

AU - D'Agostino, Giuseppe

AU - Pertwee, Roger G.

AU - MacKenzie, Alasdair

N1 - Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.

PY - 2019/11

Y1 - 2019/11

N2 - The cannabinoid-1 receptor (CB1) plays a critical role in a number of biological processes including nutrient intake, addiction and anxiety-related behaviour. Numerous studies have shown that expression of the gene encoding CB1 (CNR1) is highly dynamic with changes in the tissue specific expression of CNR1 associated with brain homeostasis and disease progression. However, little is known of the mechanisms regulating this dynamic expression. To gain a better understanding of the genomic mechanisms modulating the expression of CNR1 in health and disease we characterised the role of a highly conserved regulatory sequence (ECR1) in CNR1 intron 2 that contained a polymorphism in linkage disequilibrium with disease associated SNPs. We used CRISPR/CAS9 technology to disrupt ECR1 within the mouse genome. Disruption of ECR1 significantly reduced CNR1 expression in the hippocampus but not in the hypothalamus. These mice also displayed an altered sex-specific anxiety-related behavioural profile (open field test), reduced ethanol intake and a reduced hypothermic response following CB1 agonism. However, no significant changes in feeding patterns were detected. These data suggest that, whilst not all of the expression of CNR1 is modulated by ECR1, this highly conserved enhancer is required for appropriate physiological responses to a number of stimuli. The combination of comparative genomics and CRISPR/CAS9 disruption used in our study to determine the functional effects of genetic and epigenetic changes on the activity of tissue-specific regulatory elements at the CNR1 locus represent an important first step in gaining a mechanistic understanding of cannabinoid regulatory pharmacogenetics.

AB - The cannabinoid-1 receptor (CB1) plays a critical role in a number of biological processes including nutrient intake, addiction and anxiety-related behaviour. Numerous studies have shown that expression of the gene encoding CB1 (CNR1) is highly dynamic with changes in the tissue specific expression of CNR1 associated with brain homeostasis and disease progression. However, little is known of the mechanisms regulating this dynamic expression. To gain a better understanding of the genomic mechanisms modulating the expression of CNR1 in health and disease we characterised the role of a highly conserved regulatory sequence (ECR1) in CNR1 intron 2 that contained a polymorphism in linkage disequilibrium with disease associated SNPs. We used CRISPR/CAS9 technology to disrupt ECR1 within the mouse genome. Disruption of ECR1 significantly reduced CNR1 expression in the hippocampus but not in the hypothalamus. These mice also displayed an altered sex-specific anxiety-related behavioural profile (open field test), reduced ethanol intake and a reduced hypothermic response following CB1 agonism. However, no significant changes in feeding patterns were detected. These data suggest that, whilst not all of the expression of CNR1 is modulated by ECR1, this highly conserved enhancer is required for appropriate physiological responses to a number of stimuli. The combination of comparative genomics and CRISPR/CAS9 disruption used in our study to determine the functional effects of genetic and epigenetic changes on the activity of tissue-specific regulatory elements at the CNR1 locus represent an important first step in gaining a mechanistic understanding of cannabinoid regulatory pharmacogenetics.

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KW - enhancer

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KW - ethanol intake

KW - CB1 agonists

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KW - 212-2

KW - pharmacogenetics

KW - Ethanol intake

KW - Pharmacogenetics

KW - Enhancer

KW - NICOTINE

KW - Gene regulation

KW - Tissue specific

KW - ENDOCANNABINOID SYSTEM

KW - Anxiety-related behavior

KW - Polymorphisms

KW - HIPPOCAMPUS

KW - Wln55,212-2

KW - CB1 RECEPTORS

KW - HYPOTHALAMUS

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