Kapeleris Joanna, Kulasinghe Arutha, Warkiani Majid Ebrahimi, Oleary Connor, Vela Ian, Leo Paul, Sternes Peter, O'Byrne Kenneth, Punyadeera Chamindie
Saliva and Liquid Biopsy Translational Research Team, The School of Biomedical Sciences, Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia.
Translational Research Institute, Woolloongabba, Brisbane, Australia.
Transl Lung Cancer Res. 2020 Oct;9(5):1795-1809. doi: 10.21037/tlcr-20-521.
Tumour tissue-based information is limited. Liquid biopsy can provide valuable real-time information through circulating tumour cells (CTCs). Profiling and expanding CTCs may provide avenues to study transient metastatic disease.
Seventy non-small cell lung cancer (NSCLC) patients were recruited. CTCs were enriched using the spiral microfluidic chip and a RosetteSep™ using bloods from NSCLC patients. CTC cultures were carried out using the Clevers media under hypoxic conditions. CTCs were characterized using immunofluorescence and mutation-specific antibodies for samples with known mutation profiles. Exome sequencing was used to characterized CTC cultures.
CTCs (>2 cells) were detected in 38/70 (54.3%) of patients ranging from 0 to 385 CTCs per 7.5 mL blood. In 4/5 patients where primary tumours harboured an EGFR exon 19 deletion, this EGFR mutation was also captured in CTCs. ALK translocation was confirmed on CTCs from a patient harbouring an ALK-rearrangement in the primary tumour. Short term CTC cultures were successfully generated in 9/70 NSCLC patients. Whole exome sequencing (WES) confirmed the presence of somatic mutations in the CTC cultures with mutational signatures consistent with NSCLC.
We were able to detect CTCs in >50% of NSCLC patients. NSCLC patients with >2 CTCs had a poor prognosis. The short-term CTC culture success rate was 12.9%. Further optimization of this culture methodology may provide a means by which to expand CTCs derived from NSCLC patient's bloods. CTC cultures allow for expansion of cells to a critical mass, allowing for functional characterization of CTCs with the goal of drug sensitivity testing and the creation of CTC cell lines.
基于肿瘤组织的信息有限。液体活检可通过循环肿瘤细胞（CTC）提供有价值的实时信息。对CTC进行分析和扩增可能为研究短暂性转移性疾病提供途径。
招募了70例非小细胞肺癌（NSCLC）患者。使用螺旋微流控芯片和RosetteSep™从NSCLC患者的血液中富集CTC。在低氧条件下使用Clevers培养基进行CTC培养。使用免疫荧光和针对具有已知突变谱的样本的突变特异性抗体对CTC进行表征。外显子组测序用于表征CTC培养物。
在38/70（54.3%）的患者中检测到CTC（>2个细胞），每7.5 mL血液中CTC数量为0至385个。在5例原发肿瘤携带EGFR外显子19缺失的患者中，有4例在CTC中也检测到了该EGFR突变。在一名原发肿瘤存在ALK重排的患者的CTC中证实了ALK易位。在70例NSCLC患者中有9例成功建立了短期CTC培养物。全外显子组测序（WES）证实了CTC培养物中存在体细胞突变，其突变特征与NSCLC一致。
我们能够在超过50%的NSCLC患者中检测到CTC。CTC>2个的NSCLC患者预后较差。短期CTC培养成功率为12.9%。进一步优化这种培养方法可能提供一种扩增源自NSCLC患者血液的CTC的方法。CTC培养能够将细胞扩增到临界数量，从而实现对CTC的功能表征，目标是进行药物敏感性测试并建立CTC细胞系。
