Epigenetic regulation of F2RL3 associates with myocardial infarction and latelet function

Laura J Corbin, Stephen J White, Amy E Taylor, Christopher M Williams, Kurt Taylor, Marion T van den Bosch, Jack E Teasdale, Matthew Jones, Mark Bond, Matthew T Harper, Louise Falk, Alix Groom, Georgina G J Hazell, Lavinia Paternoster, Marcus R Munafò, Børge G Nordestgaard, Anne Tybjærg-Hansen, Stig E Bojesen, Caroline Relton, Josine L MinGeorge Davey Smith, Andrew D Mumford, Alastair W Poole, Nicholas J Timpson

Abstract

BACKGROUND: DNA hypomethylation at the F2RL3 (F2R like thrombin or trypsin receptor 3) locus has been associated with both smoking and atherosclerotic cardiovascular disease; whether these smoking-related associations form a pathway to disease is unknown. F2RL3 encodes protease-activated receptor 4, a potent thrombin receptor expressed on platelets. Given the role of thrombin in platelet activation and the role of thrombus formation in myocardial infarction, alterations to this biological pathway could be important for ischemic cardiovascular disease.

METHODS: We conducted multiple independent experiments to assess whether DNA hypomethylation at F2RL3 in response to smoking is associated with risk of myocardial infarction via changes to platelet reactivity. Using cohort data (N=3205), we explored the relationship between smoking, DNA hypomethylation at F2RL3, and myocardial infarction. We compared platelet reactivity in individuals with low versus high DNA methylation at F2RL3 (N=41). We used an in vitro model to explore the biological response of F2RL3 to cigarette smoke extract. Finally, a series of reporter constructs were used to investigate how differential methylation could impact F2RL3 gene expression.

RESULTS: Observationally, DNA methylation at F2RL3 mediated an estimated 34% of the smoking effect on increased risk of myocardial infarction. An association between methylation group (low/high) and platelet reactivity was observed in response to PAR4 (protease-activated receptor 4) stimulation. In cells, cigarette smoke extract exposure was associated with a 4.9% to 9.3% reduction in DNA methylation at F2RL3 and a corresponding 1.7-(95% CI, 1.2-2.4, P=0.04) fold increase in F2RL3 mRNA. Results from reporter assays suggest the exon 2 region of F2RL3 may help control gene expression.

CONCLUSIONS: Smoking-induced epigenetic DNA hypomethylation at F2RL3 appears to increase PAR4 expression with potential downstream consequences for platelet reactivity. Combined evidence here not only identifies F2RL3 DNA methylation as a possible contributory pathway from smoking to cardiovascular disease risk but from any feature potentially influencing F2RL3 regulation in a similar manner.

Original languageEnglish
JournalCirculation Research
Volume130
Issue number3
Pages (from-to)384-400
Number of pages17
ISSN0009-7330
DOIs
Publication statusPublished - 4 Feb 2022

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