类风湿关节炎患者外周血循环中淋巴细胞特异性蛋白酪氨酸激酶甲基化的特征研究  

Study on the characteristics of lymphocyte-specfic protein-tyrosine kinase methylation in the peripheral blood circulation of patients with rheumatoid arthritis

作　　者：许玲夏 常岑 姜平 魏凯 赵佳男 郑义馨 单雨 时一鸣 金晔华 沈逸 郭士成 何东仪 刘佳 Xu Lingxia;Chang Cen;Jiang Ping;Wei Kai;Zhao Jia′nan;Zheng Yixin;Shan Yu;Shi Yiming;Jin Ye hua;Shen Yi;Guo Shicheng;He Dongyi;Liu Jia(Department of Rheumatology,Guanghua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200052,China;Department of Medical Genetics,School of Medicine and Public Health,University of Wisconsin-Madison,Madison,WI,53706,United States)

机构地区：[1]上海中医药大学附属光华医院关节内科,上海200052 [2]美国威斯康辛大学麦迪逊分校医学与公共卫生学院医学遗传学系,美国威斯康星麦迪逊53706

出　　处：《中华风湿病学杂志》2024年第3期155-161,I0002,共8页Chinese Journal of Rheumatology

基　　金：国家自然科学基金(82074234);华东片区中西医结合关节病专科联盟项目(2021—2023);上海何东仪名医工作室建设项目(SHGZS-202220)。

摘　　要：目的分析类风湿关节炎(RA)患者外周血循环的淋巴细胞特异性蛋白酪氨酸激酶(LCK)启动子区域的甲基化特征及其与临床指标的相关性。方法采用靶向甲基化测序方法分析比较RA与健康对照(HC)和骨关节炎(OA)患者外周血LCK启动子区域7个CpG位点的甲基化水平,并与临床信息进行相关性分析和ROC曲线构建。结果采用非参数检验分析发现,与HC[0.53(0.50,0.57)]和OA患者[0.59(0.54,0.62)]相比,RA患者[0.63(0.59,0.68)]的整体甲基化水平升高(H=47.17,P<0.001)。同时与HC组[0.38(0.35,0.41)、0.59(0.55,0.63)、0.60(0.55,0.64)、0.59(0.55,0.63)、0.58(0.53,0.62)、0.45(0.43,0.49)、0.57(0.54,0.61)]相比,RA组cg0535031560、cg0535031580、cg0535031595、cg05350-315101、cg05350315104、cg05350315128和cg05350315142CpG位点的甲基化水平都显著升高[0.46(0.42,0.49)、0.70(0.65,0.75)、0.70(0.66,0.76)、0.70(0.65,0.75)、0.69(0.64,0.74)、0.55(0.51,0.59)、0.68(0.63,0.73)],差异具有统计学意义(Z=-5.63,-5.89,-5.91,-5.89,-5.98,-5.95,-5.95,均P<0.001)。与OA组相比[0.65(0.59,0.69)、0.65(0.60,0.69)、0.64(0.58,0.68)、0.50(0.45,0.54)、0.63(0.58,0.67)],RA组cg0535-031595、cg05350315101、cg05350315104、cg05350315128、cg05350315142的CpG位点甲基化水平都显著升高[0.70(0.66,0.76)、0.70(0.65,0.75)、0.69(0.64,0.74)、0.55(0.51,0.59)、0.68(0.63,0.73)],差异具有统计学意义(Z=-3.56、-3.52、-3.60、-3.67、-3.62,P=0.036、0.042、0.031、0.030、0.030)。此外,利用Pearson相关分析发现该区域整体甲基化水平与CRP(r=0.19,P=0.004)和ESR(r=0.14,P=0.035)呈正相关。CRP(低)组[0.63(0.58,0.68)]的LCK启动子区域整体甲基化水平高于CRP(高)组[0.65(0.61,0.70)],差异有统计学意义(Z=2.60,P=0.009)。最后,通过构建ROC曲线,验证外周血LCK启动子区域甲基化水平作为RA患者,特别是血清阴性RA患者与HC、OA组的鉴别效能,曲线下面积(AUC)值(95%CI)为0.78(0.63,0.93)。结论本研究提供了RA患者外周血LCK启动子区域甲基化�Objective To analyze the methylation characteristics of the lymphocyte-specific protein-tyrosine kinase(LCK)promoter region in the peripheral blood circulation of rheumatoid arthritis(RA)patients and its correlation with clinical indicators.Methods Targeted methylation sequencing was used to compare the methylation levels of 7 CpG sites in the LCK promoter region in the peripheral blood of RA patients with healthy controls(HC)and osteoarthritis(OA)patients.Correlation analysis and ROC curve construction were performed with clinical information.Results Non-parametric tests revealed that compared with HC[0.53(0.50,0.57)]and OA patients[0.59(0.54,0.62),H=47.17,P<0.001],RA patients[0.63(0.59,0.68)]exhibited an overall increase in methylation levels.Simultaneously,when compared with the HC group[0.38(0.35,0.41),0.59(0.55,0.63),0.60(0.55,0.64),0.59(0.55,0.63),0.58(0.53,0.62),0.45(0.43,0.49),0.57(0.54,0.61)],the RA group[0.46(0.42,0.49),0.70(0.65,0.75),0.70(0.66,0.76),0.70(0.65,0.75),0.69(0.64,0.74),0.55(0.51,0.59),0.68(0.63,0.73)]showed a significant elevation in methylation levels at CpG sites cg05350315_60,cg05350315_80,cg05350315_95,cg05350315_101,cg05350315_104,cg05350315_128,and cg05350315_142,with statistically significant differences(Z=-5.63,-5.89,-5.91,-5.89,-5.98,-5.95,-5.95,all P<0.001).Compared with the OA group[0.65(0.59,0.69),0.65(0.60,0.69),0.64(0.58,0.68),0.50(0.45,0.54),0.63(0.58,0.67)],the RA group[0.70(0.66,0.76),0.70(0.65,0.75),0.69(0.64,0.74),0.55(0.51,0.59),0.68(0.63,0.73)]exhibited a significant increase in methylation levels at CpG sites cg05350315_95,cg05350315_101,cg05350315_104,cg05350315_128,and cg05350315_142,with statistically significant differences(Z=-3.56,-3.52,-3.60,-3.67,-3.62;P=0.036,0.042,0.031,0.030,0.030).Furthermore,Pearson correlation coefficient analysis revealed a positive correlation between the overall methylation level in this region and C-reactive protein(CRP)(r=0.19,P=0.004)and erythrocyte sedimentation rate(r=0.14,P=0.035).The overall methylation level of the LCK promoter

关 键 词：关节炎 类风湿 甲基化 淋巴细胞特异蛋白质酪氨酸激酶p56(LCK) cg05350315 

分 类 号：R593.22[医药卫生—内科学]