马超,孙璐,赵瑜,薄剑.高通量测序结合捕获技术检测非霍奇金B细胞淋巴瘤中重链基因重排VDJ区的临床研究[J].转化医学杂志,2020,9(6):378-383
高通量测序结合捕获技术检测非霍奇金B细胞淋巴瘤中重链基因重排VDJ区的临床研究
Significance of Next Generation Sequencing for Detecting Rearranged Immunoglobulin Heavy Chain Gene in Patients with non-Hodgkin B Cell Lymphoma
  
DOI:
中文关键词:  非霍奇金B细胞淋巴瘤  免疫球蛋白重链基因  重排  高通量测序
英文关键词:Mature B cell lymphoma  Immunoglobulin heavy chain gene (IgH)  Rearrangement  Next generation sequencing (NGS)
基金项目:海南省自然科学基金(817351)
作者单位
马超 中国人民解放军总医院第一医学中心血液科 
孙璐 中国人民解放军总医院第一医学中心病理科 
赵瑜 中国人民解放军总医院第一医学中心血液科 
薄剑 中国人民解放军总医院第一医学中心血液科 
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中文摘要:
      目的探讨高通量测序(Next-Generation Sequencing,NGS)结合基因捕获技术检测非霍奇金B细胞淋巴瘤中免疫球蛋白重链基因(Immunoglobulin heavy chain gene,IgH)重排的临床应用。方法收集2019年3月至2020年5月在中国人民解放军总医院第一医学中心治疗的15例成熟B细胞淋巴瘤病例,包括11例弥漫大B细胞淋巴瘤,1例滤泡性淋巴瘤,2例边缘区淋巴瘤,1例边缘区淋巴瘤伴局部弥漫大B转化淋巴瘤。采集其初诊时的组织样本与血浆样本,通过NGS检测IgH重排,并分析其与临床病理特征的相关性。结果在组织样本中,平均检出11 212条唯一的互补决定区3(Complementarity Determining Region 3,CDR3)序列,范围从515到41 476条;在血浆样本中,平均检出1 689条唯一的CDR3序列,范围从238到6 499条。在73.3%(11/15)的患者组织中检出特异性寡克隆扩增,在其对应的血浆样本中,72.7%(8/11)存在组织中的特异性寡克隆,并且二者的比例显著相关(Spearman rho=0.803,P=0.003)。另外,存在特异性寡克隆的组织样本中,克隆指数显著高于没有特异性寡克隆的样本(P<0.05),表明存在特异性寡克隆的样本的IgH重排多样性较低。现有数据尚未发现克隆指数及组织中特异性寡克隆数与各项临床病理指标存在显著相关性。结论通过NGS检测非霍奇金B细胞淋巴患者的IgH重排,可以有效获得具体量化的CDR3序列,全面评估克隆多样性,并在大部分样本中得到特异性寡克隆序列。血浆检测结果与组织中的结果显著相关,可用于后续微小病灶残留的检测与复发监测。
英文摘要:
      ObjectiveTo explore the clinical significance of next generation sequencing (NGS) for detecting rearranged immunoglobulin heavy chain gene (IgH) in patients with mature B cell lymphoma. MethodsWe investigated 15 patients with mature B cell lymphoma treated in our hospital from March 2019 to May 2020, including 11 cases of diffuse large B cell lymphoma, one case of follicular lymphoma, two cases of marginal zone lymphoma, and one case of marginal zone lymphoma with local diffuse large B cell transformation. Rearranged IgH was detected using NGS both in tissue and plasma samples, which were collected at initial diagnosis. The relationship between IgH rearrangement and clinicopathological characteristics was also analyzed. ResultsOn average 11 212 unique complementary determining region 3 (CDR3) sequences in per tissue sample were acquired after sequencing, ranging from 515 to 41 476 sequences per sample. On average 1 689 unique CDR3 sequences per plasma sample were acquired, ranging from 238 to 6 499 sequences per sample. Specific oligoclonal expansion was detected in 73.3% (11/15) patients in the tissue samples, and 72.7% (8/11) patients have found the same specific oligoclonal expansion in their corresponding plasma samples. Furthermore, the proportion of specific oligoclone between the tissue and plasma samples is significantly correlated (Spearman rho=0.803, P=0.003). In addition, the clonality in tissue samples with specific oligoclone was significantly higher than that of samples without specific oligoclone (P<0.05), indicating that the diversity of IgH rearrangements in samples with specific oligoclone was lower. ConclusionDetection of IgH rearrangements based on NGS in patients with mature B-cell lymphoma can effectively obtain the specific quantified CDR3 sequences, comprehensively evaluate the clonal diversity, and obtain specific oligoclonal sequences in most samples. The results in plasma samples are significantly related to that in tissue samples, and could be used for minimal residual disease detection and recurrence monitoring.
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