基于正电子发射断层显像和转录组学对痰菌阴性肺结核病变组织的活动性生物标志物筛选和初步验证  

Screening and preliminary validation of biomarkers in sputum-negative pulmonary tuberculosis based on positron emission tomography/computed tomography and transcriptomics

作　　者：吴立伟 王琳[1] 温子禄 马辉[2] 欧勤芳 吴春 高欣 石磊[1] 李洪伟[1] 夏凡 宋曙 朱召芹[8] 刘海鹰[9] 陈心春 张舒林[11] 黄家颖 宋言峥[1] Wu Liwei;Wang Lin;Wen Zilu;Ma Hui;Ou Qinfang;Wu Chun;Gao Xin;Shi Lei;Li Hongwei;Xia Fan;Song Shu;Zhu Zhaoqin;Liu Haiying;Chen Xinchun;Zhang Shulin;Huang Jiaying;Song Yanzheng(Department of Thoracic Surgery,Shanghai Public Health Clinical Center,Fudan University,Shanghai 201052,China;Department of Scientific Research,Shanghai Public Health Clinical Center,Fudan University,Shanghai 201052,China;Department of Tuberculosis,Wuxi No.5 People′s Hospital,Wuxi 214007,China;Department of Tuberculosis,Changchun Infectious Disease Hospital,Changchun 130123,China;Department of PET-CT,Universal Medical Imaging,Shanghai 200233,China;Department of Lung Disease,the Navy′s 905th Hospital,Shanghai 200050,China;Department of Pathology,Shanghai Public Health Clinical Center,Fudan University,Shanghai 201052,China;Department of Laboratory,Shanghai Public Health Clinical Center,Fudan University,Shanghai 201052,China;Institute of Pathogen Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100006,China;School of Basic Medicine,Medicine Collage,Shenzhen University,Shenzhen 518060,China;School of Basic Medicine,Medicine Collage,Shanghai Jiaotong University,Shanghai 200025,China)

机构地区：[1]上海市公共卫生临床中心胸外科,上海201052 [2]上海市公共卫生临床中心科学研究部,上海201052 [3]无锡市第五人民医院呼吸科,无锡214007 [4]长春市传染病医院结核科,长春130123 [5]上海全景医学影像诊断中心,上海200233 [6]解放军海军第九O五医院肺病科,上海200050 [7]上海市公共卫生临床中心病理科,上海201052 [8]上海市公共卫生临床中心检验科,上海201052 [9]中国医学科学院北京协和医学院病原生物学研究所,北京100006 [10]深圳大学医学部基础医学院,深圳518060 [11]上海交通大学医学院基础医学院,上海200025

出　　处：《中华结核和呼吸杂志》2022年第6期567-572,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：“十三五”国家科技传染病重大专项(2017ZX10201301-003-002)。

摘　　要：目的基于正电子发射断层显像(PET-CT)和转录组学对痰菌阴性肺结核病变组织的活动性生物标志物筛选并进行初步临床验证。方法回顾性收集2017年1月1日至2019年12月31日在上海市公共卫生临床中心胸外科手术治疗的9例痰菌阴性肺结核患者作为发现组,其中男4例,女5例,年龄20~57岁,平均36岁,术前均行PET-CT扫描,术后将切除的标本按照术前PET-CT表现分为氟代脱氧葡萄糖(FDG)代谢增高区域[标准摄取值(SUV)max>3]和FDG代谢基本正常区域(SUVmax≤3)。样本处理后先对不同区域组织进行总RNA的提取,然后行全基因转录组测序。对两组数据进行生物信息学分析,发现两个区域全基因转录组数据差异的表达谱,并筛选出候选生物标志物。回顾性收集2019年1月1日至2021年1月1日上海市公共卫生临床中心收治的80例痰菌阴性肺结核患者作为验证组,其中男37例,女43例,年龄20~62岁,平均39岁,分为SUV增高组(40例)和CT影像无病灶组(40例),采用酶联免疫吸附试验(ELISA)测定患者外周血浆中候选生物标志物的蛋白水平。利用受试者工作特征(ROC)曲线对生物标志物进行效果评估。使用studentt检验来判断两组之间蛋白水平差异是否具有统计学意义。结果生物信息学分析发现,C1QB、CCL19、CCL5和HLA-DMB的表达水平与痰菌阴性肺结核病变组织的代谢活性相关。通过验证组进一步筛选验证,证实SUV增高组患者外周血浆中C1QB蛋白水平高于CT影像无病灶组,差异有统计学意义[(3.55±0.34)mg/L比(2.75±0.21)mg/L,t=4.12,P<0.001]。ROC曲线显示,C1QB蛋白水平的曲线下面积为0.731,具有潜在的临床价值。结论C1QB蛋白水平可用于评估痰菌阴性肺结核患者病变的活动性,是一种潜在的生物标志物。Objective To screen and perform preliminary clinical validation of biomarkers of activity based on positron emission tomography/computed tomography(PET-CT)and transcriptomics in sputum-negative pulmonary tuberculosis lesion tissue.Methods Nine patients with sputum-negative pulmonary tuberculosis treated surgically at the Shanghai Public Health Clinical Center for Thoracic Surgery from January 1,2017 to December 31,2019 were retrospectively collected as the discovery group,including four males and five females,aged 20-57 years(mean 36 years).All of the patients underwent PET-CT scanning before surgery,and the resected specimens were postoperatively classified according to preoperative PET-CT.The resected specimens were divided into areas with increased fluorodeoxyglucose(FDG)metabolism(SUVmax>3)and areas with normal FDG metabolism(SUVmax≤3)according to the preoperative PET-CT performance.After sample processing,total RNA was extracted from the tissues of different regions,and then whole gene transcriptome sequencing was performed.Bioinformatics analysis of the two sets of data was performed to discover the expression profiles of the differences in whole gene transcriptome data between the two regions and to screen for candidate biomarkers.Eighty patients with sputum-negative pulmonary tuberculosis admitted to Shanghai Public Health Clinical Center from January 1,2019 to January 1,2021 were retrospectively collected as the validation group,including 37 males and 43 females,aged 20-62 years,with an average age of 39 years.The validation group was divided into a group with increased SUV(n=40)and a group without lesions on CT imaging(n=40).Enzyme-linked immunosorbent assay(ELISA)was used to determine the protein levels of candidate biomarkers in the peripheral plasma of patients.The effect of biomarkers was assessed using subject operating characteristic(ROC)curves.Student′s t-test was used to determine whether the difference in protein levels between the two groups was statistically significant.Results Bioinforma

关 键 词：结核 肺 正电子发射断层显像术 生物学标记 转录组学 

分 类 号：R521[医药卫生—内科学]