Assessment of ctDNA in CSF may be a more rapid means of assessing surgical outcomes than plasma ctDNA in glioblastoma
Assessment of ctDNA in CSF may be a more rapid means of assessing surgical outcomes than plasma ctDNA in glioblastoma
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DOI：
10.1016/j.mcp.2019.06.001
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发表时间：
2019-08-01
影响因子：
3.3
通讯作者：
Mou, Yong-gao
中科院分区：
文献类型：
--
作者：
Li, Jue-hui;He, Zhen-qiang;Mou, Yong-gao
我们的目标是开发一种脑脊液 (CSF) 的高通量深度 DNA 测序分析，以确定有助于胶质母细胞瘤 (GBM) 发展的临床相关致癌突变，并作为预测患者对手术反应的生物标志物。为此，我们招募了 5 名根据术前磁共振成像诊断为高度可疑 GBM 的患者。随后，患者被组织学诊断为GBM。常规腰椎穿刺获取脑脊液，术前及术后7天采集外周血血浆。使用常规外科手术收集新鲜肿瘤样本。靶向深度测序用于表征基因组景观并识别手术前和手术后样本之间差异的突变谱。序列分析旨在检测与中枢神经系统癌症相关的 50 个基因的蛋白质编码外显子、外显子-内含子边界以及非翻译区域。从脑脊液和外周血血浆中制备循环肿瘤 DNA (ctDNA)。为了进行比较，从新鲜肿瘤组织中分离出 DNA。在脑脊液和血浆 ctDNA 中检测到非沉默编码变异，当肿瘤负荷释放（手术切除）时，前者的总体次要等位基因频率 (MAF) 与血浆相比对应于更早的疾病阶段。 GBM 的基因突变负荷与总生存期（OS，天数）显着相关（Pearson 相关性 = -0.95，P = 0.01）。我们得出的结论是，与血浆 ctDNA 相比，脑脊液 ctDNA 更好地反映了驱动基因的连续突变变化。因此，GBM 患者脑脊液的深度测序可能作为改善患者预后的替代临床检测方法。
We aimed to develop a high-throughput deep DNA sequencing assay of cerebrospinal fluid (CSF) to identify clinically relevant oncogenic mutations that contribute to the development of glioblastoma (GBM) and serve as biomarkers to predict patients' responses to surgery. For this purpose, we recruited five patients diagnosed with highly suspicious GBM according to preoperative magnet resonance imaging. Subsequently, patients were histologically diagnosed with GBM. CSF was obtained through routine lumbar puncture, and plasma from peripheral blood was collected before surgery and 7 days after. Fresh tumor samples were collected using routine surgical procedures. Targeted deep sequencing was used to characterize the genomic landscape and identify mutational profile that differed between pre-surgical and post-surgical samples. Sequence analysis was designed to detect protein-coding exons, exon-intron boundaries, and the untranslated regions of 50 genes associated with cancers of the central nervous system. Circulating tumor DNAs (ctDNAs) were prepared from the CSF and plasma from peripheral blood. For comparison, DNA was isolated from fresh tumor tissues. Non-silent coding variants were detected in CSF and plasma ctDNAs, and the overall minor allele frequency (MAF) of the former corresponded to an earlier disease stage compared with that of plasma when the tumor burden was released (surgical removal). Gene mutation loads of GBMs significantly correlated with overall survival (OS, days) (Pearson correlation = -0.95, P = 0.01). We conclude that CSF ctDNAs better reflected the sequential mutational changes of driver genes compared with those of plasma ctDNAs. Deep sequencing of the CSF of patients with GBM may therefore serve as an alternative clinical assay to improve patients' outcomes.