Disease associated polymorphisms within the conserved ECR1 enhancer differentially regulate the tissue specific activity of the cannabinoid‐1 receptor gene promoter; implications for cannabinoid pharmacogenetics

Elizabeth A Hay, Philip Cowie, Andrew R McEwan, Ruth Ross, Roger G Pertwee, Alasdair MacKenzie* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Cannabinoid receptor‐1 (CB1) represents a potential drug target against conditions that include obesity and substance abuse. However, drug trials targeting CB1 (encoded by the CNR1 gene) have been compromised by differences in patient response. Towards addressing the hypothesis that genetic changes within the regulatory regions controlling CNR1 expression contribute to these differences, we characterised the effects of disease associated allelic variation within a conserved regulatory sequence (ECR1) in CNR1 intron 2 that had previously been shown to modulate cannabinoid response, alcohol intake and anxiety‐like behaviour. We used primary cell analysis of reporters carrying different allelic variants of the human ECR1 and found that human specific C‐allele variants of ECR1 (ECR1(C)) drove higher levels of CNR1prom activity in primary hippocampal cells than did the ancestral T‐allele and demonstrated a differential response to CB1 agonism. We further demonstrate a role for the AP‐1 transcription factor in driving higher ECR1(C) activity and evidence that the ancestral t‐allele variant of ECR1 interacted with higher affinity with the insulator binding factor CTCF. The cell‐specific approaches used in our study represent an important step in gaining a mechanistic understanding the roles of non‐coding polymorphic variation in disease and in the increasingly important field of cannabinoid pharmacogenetics.

Original languageEnglish
Pages (from-to)291-298
Number of pages8
JournalHuman Mutation
Volume41
Issue number1
Early online date4 Nov 2019
DOIs
Publication statusPublished - Jan 2020

Fingerprint

Cannabinoids
Pharmacogenetics
Genes
Conserved Sequence
Nucleic Acid Regulatory Sequences
Drug Delivery Systems
Introns
Substance-Related Disorders
Transcription Factors
Obesity
Alcohols
Pharmaceutical Preparations

Keywords

  • Disease associated polymorphisms
  • Cannabinoid-1 receptor
  • gene regulation
  • tissue specific
  • promoter
  • enhancer
  • CB1 agonists
  • transcription factor
  • Chromatin immunoprecipitation
  • Cannabinoid pharmacogenetics
  • SYSTEM
  • cannabinoid pharmacogenetics
  • Win55
  • CTCF
  • FOS
  • AP-1
  • 212-2
  • chromatin immunoprecipitation
  • disease-associated polymorphisms cannabinoid-1 receptor
  • TRANSCRIPTIONAL REGULATION

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

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title = "Disease associated polymorphisms within the conserved ECR1 enhancer differentially regulate the tissue specific activity of the cannabinoid‐1 receptor gene promoter; implications for cannabinoid pharmacogenetics",
abstract = "Cannabinoid receptor‐1 (CB1) represents a potential drug target against conditions that include obesity and substance abuse. However, drug trials targeting CB1 (encoded by the CNR1 gene) have been compromised by differences in patient response. Towards addressing the hypothesis that genetic changes within the regulatory regions controlling CNR1 expression contribute to these differences, we characterised the effects of disease associated allelic variation within a conserved regulatory sequence (ECR1) in CNR1 intron 2 that had previously been shown to modulate cannabinoid response, alcohol intake and anxiety‐like behaviour. We used primary cell analysis of reporters carrying different allelic variants of the human ECR1 and found that human specific C‐allele variants of ECR1 (ECR1(C)) drove higher levels of CNR1prom activity in primary hippocampal cells than did the ancestral T‐allele and demonstrated a differential response to CB1 agonism. We further demonstrate a role for the AP‐1 transcription factor in driving higher ECR1(C) activity and evidence that the ancestral t‐allele variant of ECR1 interacted with higher affinity with the insulator binding factor CTCF. The cell‐specific approaches used in our study represent an important step in gaining a mechanistic understanding the roles of non‐coding polymorphic variation in disease and in the increasingly important field of cannabinoid pharmacogenetics.",
keywords = "Disease associated polymorphisms, Cannabinoid-1 receptor, gene regulation, tissue specific, promoter, enhancer, CB1 agonists, transcription factor, Chromatin immunoprecipitation, Cannabinoid pharmacogenetics, SYSTEM, cannabinoid pharmacogenetics, Win55, CTCF, FOS, AP-1, 212-2, chromatin immunoprecipitation, disease-associated polymorphisms cannabinoid-1 receptor, TRANSCRIPTIONAL REGULATION",
author = "Hay, {Elizabeth A} and Philip Cowie and McEwan, {Andrew R} and Ruth Ross and Pertwee, {Roger G} and Alasdair MacKenzie",
note = "EH was funded by Medical Research Scotland (PhD-719-2013) and GW Pharmaceuticals. AMcE was funded by BBSRC project grant (BB/N017544/1). PB and DW are funded by the Scottish Government Rural and Environment Science and Analytical Services Division to the Rowett Institute. The authors declare no conflicts of interest.",
year = "2020",
month = "1",
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journal = "Human Mutation",
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T1 - Disease associated polymorphisms within the conserved ECR1 enhancer differentially regulate the tissue specific activity of the cannabinoid‐1 receptor gene promoter; implications for cannabinoid pharmacogenetics

AU - Hay, Elizabeth A

AU - Cowie, Philip

AU - McEwan, Andrew R

AU - Ross, Ruth

AU - Pertwee, Roger G

AU - MacKenzie, Alasdair

N1 - EH was funded by Medical Research Scotland (PhD-719-2013) and GW Pharmaceuticals. AMcE was funded by BBSRC project grant (BB/N017544/1). PB and DW are funded by the Scottish Government Rural and Environment Science and Analytical Services Division to the Rowett Institute. The authors declare no conflicts of interest.

PY - 2020/1

Y1 - 2020/1

N2 - Cannabinoid receptor‐1 (CB1) represents a potential drug target against conditions that include obesity and substance abuse. However, drug trials targeting CB1 (encoded by the CNR1 gene) have been compromised by differences in patient response. Towards addressing the hypothesis that genetic changes within the regulatory regions controlling CNR1 expression contribute to these differences, we characterised the effects of disease associated allelic variation within a conserved regulatory sequence (ECR1) in CNR1 intron 2 that had previously been shown to modulate cannabinoid response, alcohol intake and anxiety‐like behaviour. We used primary cell analysis of reporters carrying different allelic variants of the human ECR1 and found that human specific C‐allele variants of ECR1 (ECR1(C)) drove higher levels of CNR1prom activity in primary hippocampal cells than did the ancestral T‐allele and demonstrated a differential response to CB1 agonism. We further demonstrate a role for the AP‐1 transcription factor in driving higher ECR1(C) activity and evidence that the ancestral t‐allele variant of ECR1 interacted with higher affinity with the insulator binding factor CTCF. The cell‐specific approaches used in our study represent an important step in gaining a mechanistic understanding the roles of non‐coding polymorphic variation in disease and in the increasingly important field of cannabinoid pharmacogenetics.

AB - Cannabinoid receptor‐1 (CB1) represents a potential drug target against conditions that include obesity and substance abuse. However, drug trials targeting CB1 (encoded by the CNR1 gene) have been compromised by differences in patient response. Towards addressing the hypothesis that genetic changes within the regulatory regions controlling CNR1 expression contribute to these differences, we characterised the effects of disease associated allelic variation within a conserved regulatory sequence (ECR1) in CNR1 intron 2 that had previously been shown to modulate cannabinoid response, alcohol intake and anxiety‐like behaviour. We used primary cell analysis of reporters carrying different allelic variants of the human ECR1 and found that human specific C‐allele variants of ECR1 (ECR1(C)) drove higher levels of CNR1prom activity in primary hippocampal cells than did the ancestral T‐allele and demonstrated a differential response to CB1 agonism. We further demonstrate a role for the AP‐1 transcription factor in driving higher ECR1(C) activity and evidence that the ancestral t‐allele variant of ECR1 interacted with higher affinity with the insulator binding factor CTCF. The cell‐specific approaches used in our study represent an important step in gaining a mechanistic understanding the roles of non‐coding polymorphic variation in disease and in the increasingly important field of cannabinoid pharmacogenetics.

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KW - 212-2

KW - chromatin immunoprecipitation

KW - disease-associated polymorphisms cannabinoid-1 receptor

KW - TRANSCRIPTIONAL REGULATION

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JO - Human Mutation

JF - Human Mutation

SN - 1059-7794

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