Salmon Carolin, Levermann Janina, Neves Rui P L, Liffers Sven-Thorsten, Kuhlmann Jan Dominik, Buderath Paul, Kimmig Rainer, Kasimir-Bauer Sabine
Department of Gynecology and Obstetrics, University Hospital Essen, 45147 Essen, Germany.
Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, 40225 Düsseldorf, Germany.
Cancers (Basel). 2021 Jul 26;13(15):3748. doi: 10.3390/cancers13153748.
In Ovarian Cancer (OC), the analysis of single circulating tumor cells (sCTCs) might help to investigate genetic tumor evolution during the course of treatment. Since common CTC identification features failed to reliably detect CTCs in OC, we here present a workflow for their detection and genomic analysis. Blood of 13 high-grade serous primary OC patients was analyzed, using negative immunomagnetic enrichment, followed by immunofluorescence staining and imaging for Hoechst, ERCC1, CD45, CD11b and cytokeratin (CK) and sCTC sorting with the DEPArray NxT. The whole genome of single cells was amplified and profiled for copy number variation (CNV). We detected: Type A-cells, epithelial (Hoechst, ERCC1, CD45, CD11b, CK); Type B-cells, potentially epithelial (Hoechst, ERCC1, CD45, CD11b, CK) and Type C-cells, potentially mesenchymal (Hoechst, ERCC1, CD45, CD11b, CK). In total, we identified five (38.5%) patients harboring sCTCs with an altered CN profile, which were mainly Type A-cells (80%). In addition to inter-and intra-patient genomic heterogeneity, high numbers of Type B- and C-cells were identified in every patient with their aberrant character only confirmed in 6.25% and 4.76% of cases. Further identification markers and studies in the course of treatment are under way to expand sCTC analysis for the identification of tumor evolution in OC.
在卵巢癌（OC）中，对单个循环肿瘤细胞（sCTC）进行分析可能有助于研究治疗过程中肿瘤的基因演变。由于常见的循环肿瘤细胞识别特征无法可靠地检测出OC中的循环肿瘤细胞，我们在此展示一种用于其检测和基因组分析的工作流程。对13例高级别浆液性原发性OC患者的血液进行了分析，采用阴性免疫磁珠富集法，随后进行免疫荧光染色以及对Hoechst、ERCC1、CD45、CD11b和细胞角蛋白（CK）进行成像，并使用DEPArray NxT对sCTC进行分选。对单个细胞的全基因组进行扩增，并分析其拷贝数变异（CNV）。我们检测到：A型细胞，上皮细胞（Hoechst、ERCC1、CD45、CD11b、CK）；B型细胞，可能为上皮细胞（Hoechst、ERCC1、CD45、CD11b、CK）；C型细胞，可能为间充质细胞（Hoechst、ERCC1、CD45、CD11b、CK）。总共，我们鉴定出5例（38.5%）携带拷贝数图谱改变的sCTC的患者，这些细胞主要是A型细胞（80%）。除了患者间和患者内的基因组异质性外，在每位患者中均检测到大量的B型和C型细胞，但其异常特征仅在6.25%和4.76%的病例中得到证实。目前正在进行进一步的识别标志物研究以及治疗过程中的研究，以扩展sCTC分析用于识别OC中的肿瘤演变。
