Vasantharajan Sai Shyam, Barnett Edward, Gray Elin S, McCall John L, Rodger Euan J, Eccles Michael R, Munro Fran, Pattison Sharon, Chatterjee Aniruddha
Department of Pathology, Dunedin School of Medicine, University of Otago, Dunedin 9016, New Zealand.
Centre for Precision Health and School of Medical and Health Sciences, Edith Cowan University, Joondalup, WA 6027, Australia.
Cancers (Basel). 2022 Jul 15;14(14):3446. doi: 10.3390/cancers14143446.
Circulating tumour cells (CTC) from solid tumours are a prerequisite for metastasis. Isolating CTCs and understanding their biology is essential for developing new clinical tests and precision oncology. Currently, CellSearch is the only FDA (U.S. Food and Drug Administration)-approved method for CTC enrichment but possesses several drawbacks owing to a reliance on the epithelial cell adhesion molecule (EpCAM) and a resource-intensive nature. Addressing these shortcomings, we optimised an existing size-based method, MetaCell, to enrich CTCs from blood of colorectal cancer (CRC) patients. We evaluated the ability of MetaCell to enrich CTCs by spiking blood with CRC cell lines and assessing the cell recovery rates and WBC depletion via immunostaining and gene expression. We then applied MetaCell to samples from 17 CRC patients and seven controls. Recovery rates were >85% in cell lines, with >95% depletion in WBCs. MetaCell yielded CTCs and CTC clusters in 52.9% and 23.5% of the patients, respectively, without false positives in control patients. CTCs and cluster detection did not correlate with histopathological parameters. Overall, we demonstrated that the MetaCell platform enriched CRC cells with high recovery rates and high purity. Our pilot study also demonstrated the ability of MetaCell to detect CTCs in CRC patients.
实体瘤中的循环肿瘤细胞(CTC)是转移的前提条件。分离CTC并了解其生物学特性对于开发新的临床试验和精准肿瘤学至关重要。目前,CellSearch是唯一获得美国食品药品监督管理局(FDA)批准的CTC富集方法,但由于依赖上皮细胞粘附分子(EpCAM)且资源密集,存在若干缺点。为解决这些不足,我们优化了现有的基于大小的方法MetaCell,以从结直肠癌(CRC)患者的血液中富集CTC。我们通过向血液中加入CRC细胞系,并通过免疫染色和基因表达评估细胞回收率和白细胞清除率,来评估MetaCell富集CTC的能力。然后,我们将MetaCell应用于17例CRC患者和7例对照的样本。细胞系中的回收率>85%,白细胞清除率>95%。MetaCell在52.9%的患者中产生了CTC,在23.5%的患者中产生了CTC簇,对照患者中无假阳性。CTC和簇的检测与组织病理学参数无关。总体而言,我们证明了MetaCell平台能够以高回收率和高纯度富集CRC细胞。我们的初步研究还证明了MetaCell检测CRC患者中CTC的能力。