摘要
目的利用由50个靶基因所组成的测序panel,分析骨髓增殖性肿瘤(MPN)三种主要亚型原发性血小板增多症(ET),真性红细胞增多症(PV),以及原发性骨髓纤维化(PMF)的突变谱,以便为该病的诊断和预后提供有价值的信息。方法回顾性研究方式,收集2010年至2017年间来自华山医院以及长海医院血液科的MPN患者标本共53例。所有患者的诊断均符合2016年WHO的诊断标准,包括ET31例(男11例,女20例,中位数年龄55岁),PV17例(男12例,女5例,中位数年龄65岁),PMF5例(男4例,女1例,中位数年龄67岁),并进行其骨髓或者血液标本DNA的二代测序。根据COSMIC,dbSNP,Clinvar等公共数据库对测序结果进行分析,绘制MPN人群的突变谱,并结合患者的临床随访资料进行解读。结果除去典型的JAK2、CALR、MPL突变,还检测出了另外11种基因的致病性突变,以及4种与赋予个体MPN疾病易感性相关的寡核苷酸多态性(SNP)位点(rs4858647,rs9376092,rs58270997,rs621940)。平均每个患者的突变基因数目为2.3个。全部患者(53例)中,基因突变频率由高到低依次为TET2(40%),EZH2(26%),ASXL1(11%),MIR662(6%),SF3B1(6%),BARD1(6%),DNMT3A(4%),KIT(2%),RUNX1(2%),TP53(2%),NRAS(2%)。结论利用靶向基因二代测序技术,能实现一次性对多例典型的BCR-ABL阴性的MPN患者进行多基因外显子测序,检测周期可缩短至两天,可以发现除了JAK2、CALR、MPL突变之外其他的致病性突变,在临床有较好的应用前景。
Objective By a sequencing panel consisting of 50 targeted genes, aiming at depicting the molecular landscape of ET, PV, and PMF, which are three major subtypes of MPN, to provide valuable information in the diagnosis and prognosis of MPN. Methods A retrospective study was conducted of 53 patients from Huashan hospital and Changhai hospital. All patients were diagnosed in accordance with the 2016 WHO diagnostic criteria for MPN, including 31 cases of ET(11 males, 20 females, median age 55 years), 17 cases of PV(12 males, 5 females, median age 65 years), and 5 cases of PMF(4 males, 1 females, median age 67 years), and underwent next-generation of DNA sequencing of their bone marrow or blood samples. The genetic analyses were performed on bone marrow or peripheral blood. Referring to COSMIC, dbSNP, Clinvar and other public databases, we analyzed the sequencing data, and elucidated the mutation profile of MPN patients, combining with their clinic information. Results In addition to the typical JAK2, CALR, and MPL mutations, pathogenic mutations in other 11 genes were detected, as well as 4 SNPs that confer individual susceptibility to MPNs (rs4858647, rs9376092, rs58270997, rs621940). The average rate of mutated genes was 2.3 genes per patient. In all patients (53 cases), the mutated genes detected were TET2, EZH2, ASXL1, MIR662, SF3B1, BARD1, DNMT3A, KIT, RUNX1, TP53, NRAS according to their mutational frequency. Conclusions Applying next-generation sequencing technology, multi-gene sequencing of a bunch of typical BCR-ABL-negative MPN patients can be performed at one time within 2 working days, and pathogenic mutations other than JAK2, CALR, MPL can be found, which has a bright prospection in clinic.
作者
黄秀
邓萱
许笑
张心菊
张悦晟
马玮哲
胡婷婷
唐古生
关明
Huang Xiu;Deng Xuan;Xu Xiao;Zhang Xinju;Zhang Yuesheng;Ma Weizhe;Hu Tingting;Tang Gusheng;Guan Ming(Department of Laboratory Medicine, Hua Shan Hospital, Fudan University, Shanghai 200040, China;Department of Hematology, Changhai Hospital, Naval Military Medical University, Shanghai 200433, China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2019年第6期427-434,共8页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金(81672105).
关键词
骨髓增殖性疾病
突变
高通量核昔酸序列分析
Myeloproliferative disorders
Mutation
High-throughput nucleotide sequencing
