High Detection Rate of MYD88 Mutations in Cerebrospinal Fluid From Patients With CNS Lymphomas
High Detection Rate of MYD88 Mutations in Cerebrospinal Fluid From Patients With CNS Lymphomas
复制标题
DOI：
10.1200/po.18.00308
复制
发表时间：
2019-04-18
影响因子：
4.6
通讯作者：
Fujii, Yukihiko
中科院分区：
文献类型：
--
作者：
Watanabe, Jun;Natsumeda, Manabu;Fujii, Yukihiko
活检是诊断原发性中枢神经系统淋巴瘤（PCNSL）的金标准。然而，手术活检存在与出血并发症和假阴性结果相关的发病率问题，因此需要更安全可靠的诊断方法。本研究的目的是检测MYD 88突变，一个重要的驱动突变，在脑脊液（CSF）的CNS lymphoma.PATIENTS和METHODS患者26例CNS淋巴瘤（20原发性CNS淋巴瘤和6 CNS复发的系统性淋巴瘤）进行了研究。我们通过腰椎穿刺从CSF中提取无细胞DNA（cfDNA）。从1 mL CSF中提取cfDNA，并进行桑格测序和液滴数字聚合酶链反应（ddPCR）。此外，我们对21例手术获得的福尔马林固定石蜡包埋（FFPE）组织中的MYD 88进行DNA测序，并比较结果.结果从1 mL CSF中提取的cfDNA量中位数为219 ng/mL（第25至第75百分位数，129至333 ng/mL）。CSF中MYD 88突变检出率为76.9%（20/26例），其中第5外显子L265 P突变最多，占95.0%（19/20例）。1例病例检测到第3号外显子S219 C。在4例患者中，通过ddPCR而不是通过桑格测序证实了MYD 88突变。在所有21例有足够的FFPE组织进行DNA分析的病例中，从cfDNA中检测到的MYD 88突变与从FFPE组织中检测到的肿瘤来源的DNA一致。结论该初步研究提供了证据，证明桑格测序和ddPCR相结合可以可靠地检测CNS淋巴瘤患者1 mL CSF中的cfDNA中体细胞驱动突变MYD 88。(C)2019年美国临床肿瘤学会
PURPOSE Biopsy is the gold standard for the diagnosis of primary CNS lymphoma (PCNSL). However, surgical biopsy has problems of morbidity related to hemorrhagic complications and false-negative findings, so safer and more reliable diagnostic methods are required. The aim of this study is to detect the MYD88 mutation, an important driver mutation, in the cerebrospinal fluid (CSF) of patients with CNS lymphoma.PATIENTS AND METHODS Twenty-six patients with CNS lymphoma (20 primary CNS lymphoma and six CNS relapse from systemic lymphoma) were studied. We extracted cell-free DNA (cfDNA) from CSF by lumbar puncture. cfDNA was extracted from 1 mL of CSF, and Sanger sequencing and droplet digital polymerase chain reaction (ddPCR) were performed. Furthermore, we performed DNA sequencing of MYD88 in 21 cases with available surgically obtained formalin-fixed paraffin-embedded (FFPE) tissue and compared the results.RESULTS The median cfDNA amount extracted from 1 mL CSF was 219 ng/mL (25th to 75th percentile, 129 to 333 ng/mL). MYD88 mutations were detected from CSF in 76.9% (20 of 26 cases), and L265P in exon 5 was the most frequent mutation in 19 out of 20 (95.0%) cases. S219C in exon 3 was detected in one case. In four patients, MYD88 mutation was confirmed by ddPCR but not by Sanger sequencing. In all 21 cases with sufficient FFPE tissue for DNA analysis, the detection of MYD88 mutation from cfDNA was consistent with those of tumor-derived DNA from FFPE tissue.CONCLUSION This pilot study provided evidence that the somatic driver mutation MYD88 can be reliably detected by combination of Sanger sequencing and ddPCR in the cfDNA taken from 1 mL of CSF in patients with CNS lymphomas. (C) 2019 by American Society of Clinical Oncology