Background: Multiple sclerosis (MS) is a common central nervous system autoimmune disorder. Increasing number of genome-wide association study (GWAS) analyses hint that MS is strongly associated with genetics. Unfortunately, almost all the GWAS analyses were Caucasian population based. Numbers of risk loci might not be replicated in Chinese MS patients. Hence, we performed a MassArray Assay to genotype the previously reported variants located in the transcription regulation genes in order to elucidate their role in the Chinese MS patients.
Methods: One hundred and forty-two relapsing-remitting MS (RRMS) patients and 301 healthy controls were consecutively collected from September 2, 2008, to June 7, 2013, as stage 1 subjects. Eight reported transcription regulation-related single-nucleotide polymorphisms (SNPs) were genotyped using the Sequenom MassArray system. In stage 2, another 44 RRMS patients and 200 healthy controls were consecutively collected and Sanger sequenced from April 7, 2015, to June 29, 2017, for the validation of positive results in stage 1. Differences in allele and genotype frequencies between patients and healthy controls, odds ratios, and 95% confidence intervals were calculated with the Chi-square test or Fisher's exact test. Hardy-Weinberg equilibrium was tested also using the Chi-square test.
Results: In stage 1 analysis, we confirmed only one previously reported risk variant, rs11129295 in EOMES gene. We found that the frequency of T/T genotype was much higher in MS group (χ2 = 10.251, P = 0.005) and the T allele of rs11129295 increased the risk of MS (χ2 = 10.022, P = 0.002). In stage 2 and combined analyses, the T allele of rs11129295 still increased the risk of MS (χ2 = 4.586, P = 0.030 and χ2 = 16.378, P = 5.19 × 10-5, respectively).
Conclusions: This study enhances the knowledge that the variant of EOMES is associated with increasing risk in Chinese RRMS patients and provides a potential therapeutic target in RRMS.
EOMES基因多态性与中国人群复发缓解型多发性硬化发病风险相关摘要背景：多发性硬化（Multiple Sclerosis, MS）是一种常见的中枢神经系统自身免疫性疾病。越来越多的全基因组关联分析（genome-wide association study, GWAS）表明多发性硬化可能与遗传因素密切相关。遗憾的是, 几乎所有的GWAS都是基于高加索人群， 实验中得到的很多风险位点并不能在中国MS病人身上得到验证。 方法：自2008年9月2日至2013年6月7日，我们连续收集了142例复发缓解型MS（relapsing-remitting MS, RRMS）患者及301名健康志愿者作为第一阶段的研究对象，通过Sequenom MassArray 技术对8个GWAS研究发现的转录调节相关风险位点进行单核苷酸多态性（single-nucleotide polymorphisms, SNPs）分型。为了验证第一阶段所得到的阳性结果，我们自2015年4月7日至2017年6月29日又连续收集了44例RRMS患者及200名健康志愿者，通过Sanger测序进行SNP位点检测。患者与健康志愿者基因型以及等位基因之间的差异、风险比及95%置信区间通过χ2或Fisher精确概率法进行检验。Hardy-Weinberg平衡亦通过χ2检验进行计算。 结果：在第一阶段分析中，我们发现仅有EOMES基因rs11129295位点这一个风险位点可能与MS发病风险相关，患者中携带T/T基因型的频率显著高于健康志愿者 （χ2=10.251, P=0.005），T等位基因提高MS发病风险（χ2=10.022, P= 0.002）。在第二阶段及联合分析中，rs11129295位点T等位基因依然能提高MS发病风险（χ2 = 4.586, P= 0.030 以及χ2 =16.378, P = 5.19×10-5）。 结论：本研究验证了EOMES基因多态位点rs11129295与中国RRMS患者发病风险的相关性，为新的RRMS治疗方法提供了潜在靶点。.
Keywords: Genetic Association Studies; Multiple Sclerosis; Risk Factors; Single-nucleotide Polymorphism.