Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 1L7, Canada.
Nat Commun. 2017 May 11;8:15086. doi: 10.1038/ncomms15086.
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.
骨髓抽吸物进行多发性骨髓瘤基因组分析的要求给临床试验的患者入组和保留带来了障碍。我们评估了循环无细胞 DNA (cfDNA) 分析是否可与骨髓瘤的骨髓肿瘤细胞的分子分析相媲美。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，该方法用于对 53 名骨髓瘤患者的 64 份 cfDNA 标本中的 KRAS、NRAS、BRAF、EGFR 和 PIK3CA 的所有蛋白编码外显子进行测序，中位覆盖度超过 20,000 ×。该方法包括一种变体过滤算法，可检测 cfDNA 中存在的肿瘤衍生片段，其等位基因频率低至 0.25%（中位数为 3.2%，范围为 0.25-46%）。使用 48 份 cfDNA 标本与匹配的骨髓数据进行 LB-Seq 分析，我们检测到 49/51 个可能的体细胞突变，亚克隆层次结构反映了肿瘤分析（96%的一致性），并且还有四个额外的突变可能是骨髓检测遗漏的（>98%的特异性）。总的来说，LB-Seq 是多发性骨髓瘤骨髓基因分析的一种高保真辅助手段。
Zotero 插件
