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Thromb Haemost 2014; 111(05): 960-969
DOI: 10.1160/TH13-08-0706
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Genetic analysis of the 9p21.3 CAD risk locus in Asian Indians

Jayashree Shanker   *
1   Mary and Garry Weston Functional Genomics Unit, Thrombosis Research Institute, Bangalore, India
,
Prathima Arvind   *
1   Mary and Garry Weston Functional Genomics Unit, Thrombosis Research Institute, Bangalore, India
,
Srikarthika Jambunathan
1   Mary and Garry Weston Functional Genomics Unit, Thrombosis Research Institute, Bangalore, India
,
Jiny Nair
1   Mary and Garry Weston Functional Genomics Unit, Thrombosis Research Institute, Bangalore, India
,
Vijay V. Kakkar
2   Chairman and Managing Trustee, Thrombosis Research Institute, Bangalore, India
3   Thrombosis Research Institute, London, UK
› Author Affiliations
Further Information

Publication History

Received: 29 August 2013

Accepted after major revision: 16 January 2013

Publication Date:
30 November 2017 (online)

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Summary

The 9p21.3 locus is the best replicated region to date for coronary artery disease (CAD). We investigated the association of 9p21.3 common variants with CAD, candidate gene expression including ANRIL, a non-coding RNA, followed by in vitro validation. Five variants, rs10757278, rs10757274, rs2383206, rs1333049 and rs4977574 were genotyped in 1,034 cases and 1,034 controls. Gene expression of C9orf5, MTAP 1, MTAP 2, p16INK4a , p14ARF , p15INK4b and two ANRIL splice variants, NR_003529 and EU741058, were measured in 100 cases and 100 controls. Human aortic smooth muscle cells (HuAoSMCs) were transfected with siRNA targeting ANRIL exon 19 (siRNA-1) or exon 2 (siRNA-2) and consequent effect determined. rs2383206 showed the highest association with CAD (odds ratio [OR] 2.02, 95% confidence interval [CI] 1.56 –2.62) and an adjusted OR of 2.55, 1.33–2.88 along with rs10757278. Conventional risk factors (conventional RFs), rs2383206 and rs10757278 variants together yielded a higher c index (OR 0.790, 95% CI 0.770 –0.810) as compared to conventional RFs (OR 0.783, 95% CI 0.763–0.803) or genetic variants (OR 0.561, 95% CI 0.536–0.586) alone. GAAAA haplotype showed significant protective association with CAD compared to CGGGG risk haplotype (OR 0.45, 95% CI 0.27–0.77). Expression of p16INK4a , p14ARF and p15INK4b as well as plasma CDKN2A levels were lower in cases than controls. GG genotype was associated with higher EU741058 expression and lower p16INK4a expression. HuAoSMCs transfected with siRNA-1 showed lower NR_003529, p16INK4a and p14ARF expression. Our study provides further evidence on the significance of 9p21.3 locus for CAD wherein the risk allele regulate the expression of ANRIL and adjacent tumour suppressor genes which in turn alter smooth muscle proliferation, a fundamental process in atherosclerosis.

Keywords

Coronary artery disease - 9p21.3 locus - association study - gene expression - Asian Indians

* Both authors equally contributed to the manuscript.


 

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