Schäfer Claudia, Ju Yawen, Tak Youngbin, Vazquez Cesar, Han Sangyoon J, Tan Edwin, Shay Jerry W, Holmqvist Mats, Danuser Gaudenz, Schopperle William M, Bubley Glenn
Department of Cell Biology, University of Texas, Southwestern Medical Center, Dallas, TX, USA.
CureMeta, Boston, MA, USA.
Heliyon. 2020 Jan 24;6(1):e03263. doi: 10.1016/j.heliyon.2020.e03263. eCollection 2020 Jan.
Over 90% of all cancer related deaths are due to metastasis. However, current diagnostic tools can't reliably discriminate between invasive and localized cancers.
In this proof-of-concept study, we employed the embryonic stem cell marker TRA-1-60 (TRA+) to identify TRA + cells within the blood of prostate cancer patients and searched for TRA + cells in men with metastatic and localized cancers. We isolated whole peripheral blood mononuclear cells from 26 metastatic prostate cancer patients, from 13 patients with localized prostate cancer and from 17 healthy controls. Cells were stained for DAPI, CD45 and TRA + by immunofluorescence and imaged by epi-fluorescence microscopy. Imaged-based software was used both to identify TRA + cells, and to analyze CD45 levels in TRA+ and negative cells.
We found high numbers of TRA + cells within the blood of metastatic cancer patients, whereas healthy individuals or men with localized prostate cancer showed none or very low numbers of TRA + cells. Further analysis of the CD45 levels of TRA + cells revealed a small population of TRA + cells with almost undetectable CD45 levels that were found frequently in metastatic prostate cancer patients. By excluding CD45 positive cells from the TRA + cell pool, we were able to refine the assay to be highly specific in identifying men with metastatic disease. In fact, the difference of CD45 levels between TRA+ and negative cells was a robust measure to distinguish between men with localized and metastatic prostate cancers in this small patient cohort.
The data suggest that metastatic prostate cancer patient have significant numbers of TRA+/CD45 cells which might represent a potential tool for diagnostic assessment in the future.
超过90%的癌症相关死亡是由转移所致。然而，目前的诊断工具无法可靠地区分侵袭性癌症和局限性癌症。
在这项概念验证研究中，我们采用胚胎干细胞标志物TRA-1-60（TRA+）来识别前列腺癌患者血液中的TRA+细胞，并在患有转移性和局限性癌症的男性中寻找TRA+细胞。我们从26例转移性前列腺癌患者、13例局限性前列腺癌患者和17名健康对照者中分离出全外周血单个核细胞。通过免疫荧光对细胞进行DAPI、CD45和TRA+染色，并通过落射荧光显微镜成像。基于图像的软件用于识别TRA+细胞，并分析TRA+细胞和阴性细胞中的CD45水平。
我们在转移性癌症患者的血液中发现了大量TRA+细胞，而健康个体或局限性前列腺癌男性的TRA+细胞数量为零或极少。对TRA+细胞的CD45水平进行进一步分析发现，一小部分TRA+细胞的CD45水平几乎检测不到，这在转移性前列腺癌患者中很常见。通过从TRA+细胞池中排除CD45阳性细胞，我们能够优化检测方法，使其在识别转移性疾病患者时具有高度特异性。事实上，在这个小患者队列中，TRA+细胞和阴性细胞之间CD45水平的差异是区分局限性和转移性前列腺癌男性的有力指标。
数据表明，转移性前列腺癌患者有大量TRA+/CD45细胞，这可能是未来诊断评估的一个潜在工具。
