Treatment with the GPR55 antagonist CID16020046 increases neutrophil activation in mouse atherogenesis

Thromb Haemost. 2016 Oct 28;116(5):987-997. doi: 10.1160/TH16-02-0139. Epub 2016 Jul 28.

Abstract

Endocannabinoids modulate atherogenesis by triggering different receptors. Recently, orphan G protein-coupled receptors (GPRs) were suggested to be activated by endocannabinoids, possibly regulating vasorelaxation. Here, we investigated whether GPR55 antagonism with CID16020046 would impact on atherosclerotic size and inflammation in two mouse models of early and more advanced atherogenesis. Eleven-week old ApoE-/- mice were fed either a normal diet ([ND] for 16 weeks) or a high-cholesterol diet ([HD] for 11 weeks), resulting in different degrees of hypercholesterolaemia and size of atherosclerosis. CID16020046 (0.5 mg/kg) or vehicle were intraperitoneally administrated five times per week in the last three weeks before euthanasia. Treatment with CID1602004 was well-tolerated, but failed to affect atherosclerotic plaque and necrotic core size, fibrous cap thickness, macrophage and smooth muscle cell content as well as Th cell polarisation. In ND mice, treatment with CID1602004 was associated with increased chemokine production, neutrophil and MMP-9 intraplaque content as well as reduced collagen as compared to vehicle-treated animals. In HD mice, CID1602004 increased intraplaque MMP-9 and abrogated collagen content without affecting neutrophils. In vitro, serum from CID1602004-treated ND mice increased mouse neutrophil chemotaxis towards CXCL2 as compared to serum from vehicle-treated animals. CID1602004 dose-dependently induced neutrophil degranulation that was reverted by co-incubation with the GPR55 agonist Abn-CBD. In supernatants from degranulation experiments, increased levels of the endocannabinoid and putative GPR55 ligand anandamide (AEA) were found, suggesting its possible autocrine control of neutrophil activity. These results indicate that GPR55 is critical for the negative control of neutrophil activation in different phases of atherogenesis.

Keywords: Atherosclerosis; endocannabinoids; neutrophil; receptors.

MeSH terms

  • Animals
  • Atherosclerosis / drug therapy*
  • Azabicyclo Compounds / pharmacology*
  • Benzoates / pharmacology*
  • Chemokine CXCL2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Mice
  • Mice, Knockout, ApoE
  • Neutrophil Activation*
  • Plaque, Atherosclerotic
  • Receptors, Cannabinoid*

Substances

  • 4-(4-(3-hydroxyphenyl)-3-(4-methylphenyl)-6-oxo-1H,4H,5H,6H-pyrrolo(3,4-c)pyrazol-5-yl)benzoic acid
  • Azabicyclo Compounds
  • Benzoates
  • Chemokine CXCL2
  • Cxcl2 protein, mouse
  • GPR55 protein, mouse
  • Receptors, Cannabinoid
  • Matrix Metalloproteinase 9
  • Mmp9 protein, mouse