A Combinatory Strategy for Detection of Live CTCs Using Microfiltration and a New Telomerase-Selective Adenovirus.
A Combinatory Strategy for Detection of Live CTCs Using Microfiltration and a New Telomerase-Selective Adenovirus.
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DOI：
10.1158/1535-7163.mct-14-0693
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发表时间：
2015-03
影响因子：
5.7
通讯作者：
Zhu J
中科院分区：
文献类型：
--
作者：
Ma Y;Hao S;Wang S;Zhao Y;Lim B;Lei M;Spector DJ;El-Deiry WS;Zheng SY;Zhu J
循环肿瘤细胞（CTC）已成为癌症早期诊断、预后和治疗监测的重要生物标志物。最近，由人端粒酶基因（hTERT）启动子驱动的可复制重组腺病毒显示出检测癌症患者血液样品中的活CTC。在这里，我们报告了一类新的腺病毒含有调节元件，抑制正常细胞中的hTERT基因。与仅具有hTERT核心启动子的病毒相比，新病毒在癌细胞中比在正常细胞中表现出更好的复制选择性。特别地，含有三个额外拷贝的阻遏物元件的Ad5GTSe在白细胞中显示出对癌细胞的上级向性，因此被选择用于血液样品中的CTC检测。为了进一步提高CTC鉴定的效率和特异性，我们测试了使用柔性微弹簧阵列（FMSAs）和Ad5GTSe成像的微过滤富集的组合策略。我们的实验表明，该方法有效地检测了掺入健康血液中的癌细胞以及乳腺癌和胰腺癌患者血液样品中的潜在CTC，证明了其作为检测和捕获不同肿瘤类型的CTC的高度灵敏和可靠的系统的潜力。
Circulating tumor cells (CTCs) have become an important biomarker for early cancer diagnosis, prognosis, and treatment monitoring. Recently, a replication-competent recombinant adenovirus driven by a human telomerase gene (hTERT) promoter was shown to detect live CTCs in blood samples of cancer patients. Here, we report a new class of adenoviruses containing regulatory elements that repress the hTERT gene in normal cells. Compared to the virus with only the hTERT core promoter, the new viruses showed better selectivity for replication in cancer cells than in normal cells. In particular, Ad5GTSe, containing three extra copies of a repressor element, displayed a superior tropism for cancer cells among leukocytes and was thus selected for CTC detection in blood samples. To further improve the efficiency and specificity of CTC identification, we tested a combinatory strategy of microfiltration enrichment using flexible micro spring arrays (FMSAs) and Ad5GTSe imaging. Our experiments showed that this method efficiently detected both cancer cells spiked into healthy blood and potential CTCs in blood samples of breast and pancreatic cancer patients, demonstrating its potential as a highly sensitive and reliable system for detection and capture of CTCs of different tumor types.