应用二代测序技术研究慢性淋巴细胞白血病患者的免疫球蛋白可变区特征及预后价值  

Application of next-generation sequencing technology for the investigation of immunoglobulin variable region characteristics and their prognostic significance in patients with chronic lymphocytic leukemia

作　　者：郭祯 金慧敏 邱彤璐 朱莉颖 吴雨洁 仇海荣 王琰 缪祎 金晖 范磊 李建勇 夏奕 乔纯 Guo Zhen;Jin Huimin;Qiu Tonglu;Zhu Liying;Wu Yujie;Qiu Hairong;Wang Yan;Miao Yi;Jin Hui;Fan Lei;Li Jianyong;Xia Yi;Qiao Chun(Lymphoma Center,Department of Hematology,the First Affiliated Hospital of Nanjing Medical University,Jiangsu Province Hospital,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院,江苏省人民医院血液科淋巴瘤中心,南京210029

出　　处：《中华血液学杂志》2025年第3期261-268,共8页Chinese Journal of Hematology

基　　金：国家自然科学基金(82170185、82170186);国家博士后科学基金(2021M691336);江苏省博士后科学基金(2021K083A);江苏省科教能力提升工程(ZDXK202209)。

摘　　要：目的利用二代测序(NGS)技术分析慢性淋巴细胞白血病(CLL)患者免疫球蛋白(IG)重链和轻链可变区基因特征、亚克隆情况及预后价值。方法采集2018年12月到2023年5月在江苏省人民医院初诊的36例CLL患者[包括12例B细胞受体(BCR)同型模式患者]的血液和(或)骨髓样本,应用NGS技术检测免疫球蛋白重链(IGH)、轻链(IGK、IGL)基因重排,研究IG基因在CLL中的特点及预后价值。结果NGS检测IG重链可变区(IGHV)与Sanger测序具有明显的相关性和较好的一致性(r=0.957,P<0.001)。36例患者中,9例(25.0%)患者IGHV有突变、27例(75.0%)患者IGHV无突变。MYD88突变在IGHV有突变患者中发生率更高[1/27(3.7%)对4/9(44.4%),P=0.009]。BCR同型模式IGHV#8/8B亚组患者的12号染色体三体(+12)[4/11(36.4%)对1/25(4.0%),P=0.023]的发生率更高,并更易发生Richter转化[8/11(72.7%)对4/25(16.0%),P=0.002]。36例患者使用IGKV(36/36,100%);15例使用IGLV(15/36,41.7%),其中IGLV3-21(5/15,33.3%)的使用率最高:患者均为Binet C分期及RaiⅢ~Ⅳ期,del(13)(q14)的发生率为60.0%(3/5)。使用和非使用IGLV3-21基因片段患者的中位至首次治疗时间(TTFT)分别为5.2(1.1~41.5)个月和9.9(0.1~94.4)个月。当定义2.5%的总检测片段条数(total reads)为IGHV亚克隆阈值时,检测出4例(4/36,11.1%)样本具有2个功能性克隆。有突变单克隆组(8/36,22.2%)和有突变双克隆组(1/36,2.8%)患者的中位TTFT分别为2.8(0.9~72.7)个月和12.8个月,中位总生存(OS)期分别为57.5(32.0~120.7)个月和51.8个月;无突变单克隆组(24/36,66.7%)和无突变双克隆组(2/36,5.6%)患者的中位TTFT分别为10.9(0.3~94.4)个月和6.3(0.1~12.5)个月,中位OS期分别为49.9(22.2~211.1)个月和30.0(9.6~50.3)个月(P>0.05)。结论应用NGS技术检测IG基因重排,不仅能分析IGHV的突变状态、优势克隆、判断预后,还能够研究IGK/IGL的基因重排片段及亚克隆的使用情况,为IGLV3-21 CLL的不良预后研究提供了线索。IGHV无突�ObjectiveTo elucidate the genomic characteristics of the immunoglobulin(IG)heavy-chain variable region and light-chain variable region,the expression of subclones,and the prognostic significance in patients with CLL.MethodsBlood and/or bone marrow specimens were gathered from a cohort of 36 patients with CLL diagnosed at Jiangsu Province Hospital from December 2018 to May 2023,including 12 cases of B cell receptor(BCR)stereotyped patients.IG heavy-chain(IGH)and light-chain(IG Kappa[IGK]and IG lambda[IGL])gene rearrangements were performed using next-generation sequencing(NGS)technology to analyze the characteristics and prognostic value in CLL.ResultsNGS detection of IG variable region(IGHV)demonstrated a significant correlation and superior consistency with Sanger sequencing(r=0.957,P<0.001).Among the 36 patients,the IGH variant(IGHV)was observed in 9(25.0%)but not in 27(75.0%)participants.The incidence of the MYD88 mutation was higher among patients with mutated IGHV[1/27(3.7%)vs 4/9(44.4%),P=0.00].A high incidence of trisomy 12 was observed in the IGHV#8/#8B subset[4/11(36.4%)vs 1/25(4.0%),P=0.023],which were more likely to develop Richter transformation[8/11(72.7%)vs 4/25(16.0%),P=0.002].In the patient cohort,36 individuals(36/36,100.0%)used the IGK variable,whereas 15 individuals(15/36,41.7%)employed the IGL variable(IGLV).IGLV3-21 reported the highest utilization rate in IGLV(5/15,33.3%).Remarkably,patients with CLL with IGLV3-21 fragments were exclusively observed in the Binet C stage and Rai PhaseⅢ-Ⅳ,with an incidence of del(13)(q14)at 60.0%(3/5).The median time to first treatment(TTFT)of patients with or without IGLV3-21 fragments was 5.2(1.1-41.5)and 9.9(0.1-94.4)months,respectively.Using the total reads threshold of 2.5%,4(4/36,11.1%)samples were detected to have two IGHV productive clones.The median TTFT and overall survival(OS)time were 2.8(0.9-72.7)and 12.8 months in patients with one mutated clone and 57.5(32.0-120.7)and 51.8 months in those with two mutated clones,respectively.The median TTFT

关 键 词：白血病 淋巴细胞 慢性 B细胞 免疫球蛋白 二代测序 亚克隆 IGLV3-21 

分 类 号：R733.7[医药卫生—肿瘤]