Liu Haiyan E, Vuppalapaty Meghah, Wilkerson Charles, Renier Corinne, Chiu Michael, Lemaire Clementine, Che James, Matsumoto Melissa, Carroll James, Crouse Steve, Hanft Violet R, Jeffrey Stefanie S, Di Carlo Dino, Garon Edward B, Goldman Jonathan, Sollier Elodie
Vortex Biosciences, Inc., Pleasanton, CA, United States.
Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, United States.
Front Oncol. 2020 Oct 8;10:572895. doi: 10.3389/fonc.2020.572895. eCollection 2020.
Lung cancer is the leading cause of cancer-related mortality worldwide. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) therapies, based on the evaluation of mutations, have shown dramatic clinical benefits. mutation assays are mainly performed on tumor biopsies, which carry risks, are not always successful and give results relevant to the timepoint of the assay. To detect secondary mutations, which cause resistance to 1st and 2nd generation TKIs and lead to the administration of a 3rd generation drug, effective and non-invasive monitoring of mutation status is needed. Liquid biopsy analytes, such as circulating tumor cells (CTCs) and circulating tumor DNA (cfDNA), allow such monitoring over the course of the therapy. The aim of this study was to develop and optimize a workflow for the evaluation of cfDNA and CTCs in NSCLC patients all from one blood sample. Using Vortex technology and EntroGen ctEGFR assay, mutations were identified at 0.5 ng of DNA (∼83 cells), with a sensitivity ranging from 0.1 to 2.0% for a total DNA varying from 25 ng (∼4 CTCs among 4000 white blood cells, WBCs) to 1 ng (∼4 CTCs among 200 WBCs). The processing of plasma-depleted-blood provided comparable capture recovery as whole blood, confirming the possibility of a multimodality liquid biopsy analysis (cfDNA and CTC DNA) from a single tube of blood. Different anticoagulants were evaluated and compared in terms of respective performance. Blood samples from 24 NSCLC patients and 6 age-matched healthy donors were analyzed with this combined workflow to minimize blood volume needed and sample-to-sample bias, and the mutation profile detected from CTCs and cfDNA was compared to matched tumor tissues. Despite the limited size of the patient cohort, results from this non-invasive mutation analysis are encouraging and this combined workflow represents a valuable means for informing therapy selection and for monitoring treatment of patients with NSCLC.
肺癌是全球癌症相关死亡的主要原因。基于突变评估的表皮生长因子受体（EGFR）酪氨酸激酶抑制剂（TKI）疗法已显示出显著的临床益处。突变检测主要在肿瘤活检组织上进行，存在风险，并不总是成功，且给出的结果与检测时间点相关。为了检测导致对第一代和第二代TKI产生耐药性并促使使用第三代药物的继发性突变，需要对突变状态进行有效且非侵入性的监测。液体活检分析物，如循环肿瘤细胞（CTC）和循环肿瘤DNA（cfDNA），可在治疗过程中进行此类监测。本研究的目的是开发并优化一种工作流程，用于评估来自同一血样的非小细胞肺癌（NSCLC）患者的cfDNA和CTC。使用涡旋技术和EntroGen ctEGFR检测方法，在0.5 ng DNA（约83个细胞）时可鉴定出突变，对于总量从25 ng（4000个白细胞中约4个CTC）到1 ng（200个白细胞中约4个CTC）的DNA，灵敏度范围为0.1%至2.0%。去除血浆的血液处理提供了与全血相当的捕获回收率，证实了从单管血液进行多模态液体活检分析（cfDNA和CTC DNA）的可能性。对不同抗凝剂的各自性能进行了评估和比较。使用这种组合工作流程分析了24例NSCLC患者和6例年龄匹配的健康供体的血样，以尽量减少所需血量和样本间偏差，并将从CTC和cfDNA检测到的突变谱与匹配的肿瘤组织进行比较。尽管患者队列规模有限，但这种非侵入性突变分析的结果令人鼓舞，并且这种组合工作流程是指导NSCLC患者治疗选择和监测治疗的有价值手段。
