Circulating Tumor Cell Detection and Capture by Photoacoustic Flow Cytometry in Vivo and ex Vivo.
Circulating Tumor Cell Detection and Capture by Photoacoustic Flow Cytometry in Vivo and ex Vivo.
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DOI：
10.3390/cancers5041691
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发表时间：
2013-12-10
期刊：
影响因子：
5.2
通讯作者：
Zharov VP
中科院分区：
文献类型：
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作者：
Galanzha EI;Zharov VP
尽管在检测循环肿瘤细胞(CTCs)方面取得了进展，但现有的检测方法由于血样体积较小(5-10毫升)，灵敏度仍然较低(1-10毫升/毫升)。因此，它们可能会漏掉多达103-104个CTC，导致勉强可以治疗的转移。在这里，我们分析了一种通过检测体内全血容量(成人为5 L)来提高灵敏度(高达102-103倍)的体内CTC检测的新概念。我们的重点是利用无标记或靶向检测的体内光声(PA)流式细胞术(PAFC)、具有超强PA共振的可光开关纳米颗粒、光纤-磁性PA探针的磁性捕获、光学清除、实时光谱识别、非线性信号放大，以及与PAFC的体外整合。我们展示了PAFC在临床前动物模型中检测罕见的白血病、鳞癌、黑色素瘤和乳腺块状和干性CTCs及其簇的能力，以及触诊、活检或手术引发的原发肿瘤中CTC的释放，从而增加转移风险。根据CTC转移潜能的不同，CTC的寿命在0.5-4h之间，这是血管内和外渗速率之间的平衡。我们介绍了作为纳米气泡增强的PA诊断、光热治疗和通过CTC计数进行反馈的CTC的声学疗法。体内数据与体外PAFC验证表明，与常规检测方法相比，该方法具有更高的灵敏度(1CTC/40毫升)和吞吐量(最高可达10毫升/分钟)。进一步的发展包括检测与癌症相关的循环微粒，以及超越衍射和光谱限制的超分辨率PAFC。
Despite progress in detecting circulating tumor cells (CTCs), existing assays still have low sensitivity (1–10 CTC/mL) due to the small volume of blood samples (5–10 mL). Consequently, they can miss up to 103–104 CTCs, resulting in the development of barely treatable metastasis. Here we analyze a new concept of in vivo CTC detection with enhanced sensitivity (up to 102–103 times) by the examination of the entire blood volume in vivo (5 L in adults). We focus on in vivo photoacoustic (PA) flow cytometry (PAFC) of CTCs using label-free or targeted detection, photoswitchable nanoparticles with ultrasharp PA resonances, magnetic trapping with fiber-magnetic-PA probes, optical clearance, real-time spectral identification, nonlinear signal amplification, and the integration with PAFC in vitro. We demonstrate PAFC’s capability to detect rare leukemia, squamous carcinoma, melanoma, and bulk and stem breast CTCs and its clusters in preclinical animal models in blood, lymph, bone, and cerebrospinal fluid, as well as the release of CTCs from primary tumors triggered by palpation, biopsy or surgery, increasing the risk of metastasis. CTC lifetime as a balance between intravasation and extravasation rates was in the range of 0.5–4 h depending on a CTC metastatic potential. We introduced theranostics of CTCs as an integration of nanobubble-enhanced PA diagnosis, photothermal therapy, and feedback through CTC counting. In vivo data were verified with in vitro PAFC demonstrating a higher sensitivity (1 CTC/40 mL) and throughput (up to 10 mL/min) than conventional assays. Further developments include detection of circulating cancer-associated microparticles, and super-resolution PAFC beyond the diffraction and spectral limits.