Department of Medicine, University of Southern California (USC), Keck School of Medicine and Norris Comprehensive Cancer Center (NCCC), California, Los Angeles.
Department of Translational Genomics, USC Keck School of Medicine and NCCC, California, Los Angeles.
Int J Cancer. 2020 May 15;146(10):2845-2854. doi: 10.1002/ijc.32915. Epub 2020 Feb 27.
Transcriptomic profiling of metastatic cancer can illuminate mechanisms of progression and lead to new therapies, but standard biopsy is invasive and reflects only a single metastatic site. In contrast, circulating tumor cell (CTC) profiling is noninvasive and repeatable, reflecting the dynamic and systemic nature of advanced disease. To date, transcriptomic profiling of CTCs has not delivered on its full potential, because white blood cells (WBCs) vastly outnumber CTCs. Current profiling strategies either lack cancer sensitivity and specificity or require specialized CTC capture protocols that are not readily scalable to large patient cohorts. Here, we describe a new strategy for rapid CTC enrichment and transcriptomic profiling using commercially available WBC depletion, microfluidic enrichment and RNA sequencing. When applied to blood samples from patients with advanced prostate cancer (PC), transcriptomes from enriched samples cluster with cancer positive controls and previously undetectable prostate-specific transcripts become readily measurable. Gene set enrichment analysis reveals multiple significantly enriched signaling pathways associated with PC, as well as novel pathways that merit further study. This accessible and scalable approach yields cancer-specific transcriptomic data and can be applied repeatedly and noninvasively in large cancer patient cohorts to discover new therapeutic targets in advanced disease.
转移性癌症的转录组谱分析可以阐明进展的机制,并导致新的治疗方法,但标准活检具有侵入性,只能反映单个转移性部位。相比之下,循环肿瘤细胞(CTC)的分析是无创的,且可以重复,反映了晚期疾病的动态和系统性。迄今为止,CTC 的转录组谱分析尚未充分发挥其潜力,因为白细胞(WBC)的数量远远超过 CTC。目前的分析策略要么缺乏癌症的敏感性和特异性,要么需要专门的 CTC 捕获方案,这些方案不容易扩展到大患者队列。在这里,我们描述了一种使用商业上可用的白细胞去除、微流控富集和 RNA 测序进行快速 CTC 富集和转录组分析的新策略。当应用于晚期前列腺癌(PC)患者的血液样本时,富集样本的转录组与阳性对照的癌症样本聚类,以前无法检测到的前列腺特异性转录本变得易于测量。基因集富集分析揭示了与 PC 相关的多个显著富集的信号通路,以及值得进一步研究的新通路。这种易于使用且可扩展的方法可产生癌症特异性的转录组数据,并可在大型癌症患者队列中反复进行无创检测,以发现晚期疾病中的新治疗靶点。