RAS/RAF mutations in tumor samples and cell-free DNA from plasma and bone marrow aspirates in multiple myeloma patients
RAS/RAF mutations in tumor samples and cell-free DNA from plasma and bone marrow aspirates in multiple myeloma patients
复制标题
多发性骨髓瘤患者肿瘤样本以及血浆和骨髓抽吸物中的游离 DNA 中的 RAS/RAF 突变
DOI：
10.7150/jca.43729
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发表时间：
2020-01-01
影响因子：
3.9
通讯作者：
Liu, Zhiqiang
中科院分区：
文献类型：
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作者：
Li, Qian;Huang, Helen J.;Liu, Zhiqiang
用途：为了评估多发性骨髓瘤（MM）骨髓活检和血浆循环游离DNA（cfDNA）中基因突变的检测。实验设计：我们使用来自血浆和骨髓的游离DNA，使用液滴数字PCR（ddPCR）的多重测定来测试BRAF V600、KRAS G12/G13、NRAS G12/G13和NRAS Q61突变，并评估结果与临床结局。结果：在83例患者中，骨髓中上述4个基因的突变频率分别为4（5%）、13（16%）、3（4%）和14（17%）。大多数突变的中位变异等位基因频率（VAF）为1.595%。在17对cfDNA样本中，上述4个基因的可检测突变频率分别为5（30%）、1（6%）、0（0%）和3（18%），中位VAF率为2.9%。与组织检测相比，骨髓DNA和血浆cfDNA之间的一致性分别为76%、100%、100%和100%。在17例骨髓和血浆样本配对的患者中，上述4种突变分别为3（18%）、1（6%）、0（0%）和2（12%），与组织检测的符合率分别为88%、88%、100%和100%。结论：ddPCR检测骨髓和血浆cfDNA中BRAF、KRAS和NRAS基因突变，高突变VAF患者生存期短。
Purpose: To evaluate the detection of gene mutations in bone marrow biopsy and circulating free DNA (cfDNA) from plasma in multiple myeloma (MM).Experimental design: We used cell-free DNA from plasma and bone marrow to test BRAF V600, KRAS G12/G13, NRAS G12/G13 and NRAS Q61 mutations using multiplex assays for droplet digital PCR (ddPCR), and evaluated results with clinical outcomes.Results: We found of 83 patients, the detectable mutation frequencies for the above four genes were 4 (5%), 13 (16%), 3 (4%) and 14 (17%) in bone marrow, respectively. The median variant allelic frequency (VAF) in most mutations were 1.595%. In 17 paired cfDNA samples, the detectable mutation frequencies for the above four genes were 5 (30%), 1 (6%), 0 (0%) and 3 (18%) respectively, and the median VAF rate was 2.9%. Agreement between bone marrow DNA and plasma cfDNA were 76%, 100%, 100% and 100% compared to the tissue detections, respectively. In 17 patients with paired bone marrow and plasma samples, the above four mutations were 3 (18%), 1 (6%), 0 (0%) and 2 (12%) respectively, with the agreement rates of 88%, 88%, 100% and 100% compared to tissue detections. Of 57 patients with available outcome data, high mutation VAF had a shorter median survival than patients with low mutation VAF (P=0.0322).Conclusions: Oncogenic mutations in BRAF, KRAS and NRAS genes can be detected in the bone marrow and plasma cfDNA with ddPCR in patients with MM patients and high VAF is associated with short survival.