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Influence of a Coronary Artery Disease-Associated Genetic Variant on FURIN Expression and Effect of Furin on Macrophage Behavior.

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journal contribution
posted on 09.08.2018, 14:21 by Guojun Zhao, Wei Yang, Jingchun Wu, Bairu Chen, Xu Yang, Junhui Chen, David G. McVey, Catherine Andreadi, Peng Gong, Tom R. Webb, Nilesh J. Samani, Shu Ye
OBJECTIVE: Genome-wide association studies have revealed a robust association between genetic variation on chromosome 15q26.1 and coronary artery disease (CAD) susceptibility; however, the underlying biological mechanism is still unknown. The lead CAD-associated genetic variant (rs17514846) at this locus resides in the FURIN gene. In advanced atherosclerotic plaques, furin is expressed primarily in macrophages. We investigated whether this CAD-associated variant alters FURIN expression and whether furin affects monocyte/macrophage behavior. APPROACH AND RESULTS: A quantitative reverse transcription polymerase chain reaction analysis showed that leukocytes from individuals carrying the CAD risk allele of rs17514846 had increased FURIN expression. A chromatin immunoprecipitation assay revealed higher RNA polymerase II occupancy in the FURIN gene in mononuclear cells of individuals carrying this allele. A reporter gene assay in transiently transfected monocytes/macrophages indicated that the CAD risk allele had higher transcriptional activity than the nonrisk allele. An analysis of isogenic monocyte cell lines created by CRISPR-mediated genome editing showed that isogenic cells with the A/A genotype for rs17514846 had higher FURIN expression levels than the isogenic cells with the C/C genotype. An electrophoretic mobility shift assay exhibited preferential binding of a nuclear protein to the risk allele. Studies of monocytes/macrophages with lentivirus-mediated furin overexpression or shRNA-induced furin knockdown showed that furin overexpression promoted monocyte/macrophage migration, increased proliferation, and reduced apoptosis whereas furin knockdown had the opposite effects. CONCLUSIONS: Our study shows that the CAD-associated genetic variant increases FURIN expression and that furin promotes monocyte/macrophage migration and proliferation while inhibiting apoptosis, providing a biological mechanism for the association between variation at the chromosome 15q26.1 locus and CAD risk.

Funding

We are thankful for the support of the British Heart Foundation (PG/16/9/31995, RG/16/13/32609), National Natural Science Foundation of China (81370202), Guangxi Natural Sciences Foundation (AA139209), and China Postdoctoral Science Foundation (2015M572342). This work falls under the portfolio of research conducted within the National Institute for Health Research Leicester Biomedical Research Centre.

History

Citation

Arteriosclerosis, Thrombosis, and Vascular Biology. 2018;38:1837-1844

Author affiliation

/Organisation/COLLEGE OF LIFE SCIENCES/School of Medicine/Department of Cardiovascular Sciences

Version

VoR (Version of Record)

Published in

Arteriosclerosis

Publisher

American Heart Association, Lippincott, Williams & Wilkins

issn

1079-5642

eissn

1524-4636

Acceptance date

13/06/2018

Copyright date

2018

Available date

09/08/2018

Publisher version

https://www.ahajournals.org/doi/10.1161/ATVBAHA.118.311030

Notes

The online-only Data Supplement is available with this article at https://www.ahajournals.org/journal/atvb/doi/suppl/10.1161/atvbaha.118.311030.

Language

en