Circulating tumour DNA sequence analysis as an alternative to multiple myeloma bone marrow aspirates.
Circulating tumour DNA sequence analysis as an alternative to multiple myeloma bone marrow aspirates.
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DOI：
10.1038/ncomms15086
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发表时间：
2017-05-11
影响因子：
16.6
通讯作者：
Pugh TJ
中科院分区：
文献类型：
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作者：
Kis O;Kaedbey R;Chow S;Danesh A;Dowar M;Li T;Li Z;Liu J;Mansour M;Masih-Khan E;Zhang T;Bratman SV;Oza AM;Kamel-Reid S;Trudel S;Pugh TJ
多发性骨髓瘤基因组分析需要骨髓穿刺液，这对临床试验中患者的招募和保留构成了障碍。我们评估了循环无细胞DNA（cfDNA）分析是否与使用骨髓肿瘤细胞的骨髓瘤分子谱分析相当。我们在此报告了一种基于杂交捕获的液体活检测序（LB-Seq）方法，用于对来自53名骨髓瘤患者的64份cfDNA标本中KRAS、NRAS、BRAF、EGFR和PIK 3CA的所有蛋白编码外显子进行测序，覆盖率> 20，000 ×中位数。该方法包括变体过滤算法，其能够检测cfDNA中存在的等位基因频率低至0.25%（中值3.2%，范围0.25-46%）的肿瘤衍生片段。使用具有匹配骨髓数据的48个cfDNA样本的LB-Seq分析，我们检测到49/51个可能的体细胞突变，其中亚克隆层次反映了肿瘤谱（96%一致性），并且骨髓检测可能遗漏了另外4个突变（>98%特异性）。总体而言，LB-Seq是多发性骨髓瘤骨髓遗传图谱的高保真辅助工具。多发性骨髓瘤的基因分析需要进行痛苦的骨髓活检。在这里，作者开发了一种替代的非侵入性方法，用于对骨髓瘤患者循环无细胞DNA中的五种癌基因进行测序，证明与骨髓肿瘤分析结果的一致性为96%。
The requirement for bone-marrow aspirates for genomic profiling of multiple myeloma poses an obstacle to enrolment and retention of patients in clinical trials. We evaluated whether circulating cell-free DNA (cfDNA) analysis is comparable to molecular profiling of myeloma using bone-marrow tumour cells. We report here a hybrid-capture-based Liquid Biopsy Sequencing (LB-Seq) method used to sequence all protein-coding exons of KRAS, NRAS, BRAF, EGFR and PIK3CA in 64 cfDNA specimens from 53 myeloma patients to >20,000 × median coverage. This method includes a variant filtering algorithm that enables detection of tumour-derived fragments present in cfDNA at allele frequencies as low as 0.25% (median 3.2%, range 0.25–46%). Using LB-Seq analysis of 48 cfDNA specimens with matched bone-marrow data, we detect 49/51 likely somatic mutations, with subclonal hierarchies reflecting tumour profiling (96% concordance), and four additional mutations likely missed by bone-marrow testing (>98% specificity). Overall, LB-Seq is a high fidelity adjunct to genetic profiling of bone-marrow in multiple myeloma. Genetic profiling of multiple myeloma requires painful bone marrow biopsies. Here, the authors develop an alternative non-invasive method for sequencing of five oncogenes in circulating cell-free DNA from myeloma patients, demonstrating 96% concordance with bone marrow tumour profiling results.