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Two-stage association study in Chinese Han identifies two independent associations in CCR1/CCR3 locus as candidate for Behçet’s disease susceptibility

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Abstract

Previous GWAS studies from Turkey suggested a potential risk locus at CCR1/CCR3 for Behçet’s disease. However, this locus did not reach the GWAS significance threshold and has not yet been examined in other ethnic populations. The current study aimed to explore whether this locus was associated with Behçet’s disease in Chinese Han and the functional role of the identified variants. A two-stage association study was performed in 653 patients and 1,685 controls using the iPLEX system. Real-time PCR was performed to examine the expression level of CCR1 and CCR3 genes. Haplotype analysis was used to construct the haplotype block. Logistic regression analysis was applied to calculate the independence of multiple associations. Bonferroni correction was applied to account for multiple testing. First stage analysis showed that ten SNPs, located in 3′UTR, 5′UTR in CCR1 or 5′UTR in CCR3, were significantly associated with Behçet’s disease (P c = 0.018 to 1.3 × 10−3). The associations of six SNPs within this locus are independent after control for the genetic effect of rs17282391 using logistic regression analysis. Haplotype analysis identified three associated haplotypes: H3 (GTGAC), H6 (CCATTA) and H9 (CGA) (P c = 0.04 to 7.79 × 10−4). Three SNPs rs13084057, rs13092160 and rs13075270 showed consistent association in replication and combining studies (replication P c = 5.31 × 10−5 to 1.44 × 10−5; combining P c = 2.76 × 10−7 to 6.50 × 10−8). Interestingly, eQTLs database reveals that SNP rs13092160 is eQTLs SNP, suggesting that this SNP is likely to be functional SNP that directly affects gene expression. The expression of CCR1 and CCR3 was increased in individuals with the CT genotype of rs13092160 (P < 0.05). No significant difference was found for the mRNA level of CCR1 and CCR3 between Behçet’s patients and controls. These findings strongly indicate CCR1/CCR3 as a novel locus underlying Behçet’s disease.

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Abbreviations

GWAS:

Genome-wide association study

HLA :

Human leukocyte antigen

SNP:

Single nucleotide polymorphism

OR:

Odds ratio

95 % CI:

95 % confidence interval

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Acknowledgments

The samples of patients and healthy controls were partially collected in the Zhongshan Ophthalmic center, Sun Yat-sen University. We thank Ms Hongyan Zhou and Mr Xiangkun Huang for their assistance in sample collection and technical support. We thank all donors enrolled in the present study. This work was supported by Natural Science Foundation Major International (Regional) Joint Research Project (30910103912), National Natural Science Foundation Project (81070723), Program for the Training of a Hundred Outstanding S&T Leaders of Chongqing Municipality, Key Project of Health Bureau of Chongqing (2012-1-003), Chongqing Key Laboratory of Ophthalmology (CSTC, 2008CA5003), Key Project of Natural Science Foundation (81130019), Research Fund for the Doctoral Program of Higher Education of China (20115503110002) and Fund for PAR-EU Scholars Program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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None of the authors has a proprietary or financial interest in any of the products mentioned.

Ethical standard

This study was approved by the Clinical Research Ethics Committee of the First Affiliated Hospital of Chongqing Medical University and Zhongshan Ophthalmic Center, Sun Yat-sen University.

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Correspondence to Peizeng Yang.

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Hou, S., Xiao, X., Li, F. et al. Two-stage association study in Chinese Han identifies two independent associations in CCR1/CCR3 locus as candidate for Behçet’s disease susceptibility. Hum Genet 131, 1841–1850 (2012). https://doi.org/10.1007/s00439-012-1200-4

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