Polymer nanofiber-embedded microchips for detection, isolation, and molecular analysis of single circulating melanoma cells.
Polymer nanofiber-embedded microchips for detection, isolation, and molecular analysis of single circulating melanoma cells.
复制标题
DOI：
10.1002/anie.201208452
复制
发表时间：
2013-03-18
影响因子：
16.6
通讯作者：
Tseng, Hsian-Rong
中科院分区：
文献类型：
--
作者：
Hou, Shuang;Zhao, Libo;Shen, Qinglin;Yu, Juehua;Ng, Charles;Kong, Xiangju;Wu, Dongxia;Song, Min;Shi, Xiaohong;Xu, Xiaochun;OuYang, Wei-Han;He, Rongxian;Zhao, Xing-Zhong;Lee, Tom;Brunicardi, F. Charles;Garcia, Mitch Andre;Ribas, Antoni;Lo, Roger S.;Tseng, Hsian-Rong
循环肿瘤细胞（CTC）[1]是从原发性肿瘤或转移部位脱落的癌细胞。外周血中CTC的存在和数量可以分别提供关于预后和治疗反应模式的临床重要数据。[2]因此，与能够进行金标准病理学分析的传统侵入性肿瘤活检一样，CTC可以被认为是肿瘤的“液体活检”，其能够重复且相对非侵入性地表征肿瘤演变，这在治疗干预期间尤其重要。目前，FDA批准的CellSearch测定法在捕获CTC方面是昂贵且低效的，并且富集的CTC通常被大量的白色血细胞（WBC）污染。因此，CTC的诊断价值未得到充分利用。在过去的十年中，已经开发了多种CTC检测方法[2d，3]，以克服基于免疫磁性分离的CellSearch测定所遇到的挑战。与现有的CTC诊断方法不同，[2d，3]我们已经证明了“纳米维可牢”芯片[4]，其能够以极高的效率富集、识别和计数患者血液样品中的CTC。首先，我们开创了一种独特的nanovelcro基底概念，[5]通过该概念，抗EpCAM涂层的硅纳米线（SiNW）基底被用于在固定设备中固定CTC。我们最近表明，其他类型的纳米结构基底（例如，电化学沉积的共轭聚合物纳米特征[6]和水平堆积的TiO 2纳米纤维[7]）也表现出增强的捕获CTCs的亲和力，证明了纳米维可牢尼龙搭扣基底的普遍适用性。我们的方法是独特的，因为使用了纳米结构的基底;在抗EpCAM包被的纳米基底和CTC上的纳米尺度细胞表面组分（例如，微绒毛）之间存在增强的局部地形相互作用[8]，这类似于维可牢尼龙搭扣的工作原理。其次，通过将光刻图案化的纳米维可牢基底与覆盖的聚二甲基硅氧烷（PDMS）混沌混合器[9]集成，提高了流经系统和基底的CTC之间的接触频率，我们进一步提高了CTC捕获效率。[4]使用人工和患者CTC样本的并行分析验证研究表明，nanovelcro芯片的灵敏度优于CellSearch。尽管nanovelcro芯片允许有效且可再现地检测患者血液中的CTC，但挑战仍然存在于1）扩大其用于检测表现出除EpCAM之外的表面标志物的其他类型的实体瘤CTC的一般适用性，以及2）使得能够分离单个CTC用于随后的分子分析。为了拓宽基于纳米velcro的细胞亲和性测定的一般适用性，我们探索了一种黑色素瘤特异性捕获剂[13]（即抗CD 146），以捕获循环黑色素瘤细胞（CMC;实体瘤CTC的一个亚类）。此外，
Circulating tumor cells (CTCs)[1] are cancer cells shed from either primary tumors or metastatic sites. The presence and number of CTCs in peripheral blood can provide clinically important data on prognosis and therapeutic response patterns, respectively.[2] Thus, as with traditional invasive tumor biopsies that enable gold-standard pathological analysis, CTCs can be regarded as “liquid biopsies” of the tumor, which enable repeated and relatively non-invasive characterization of tumor evolution, especially important during therapeutic interventions. Currently, the FDA-approved CellSearch assay is costly and inefficient at capturing CTCs, and the enriched CTCs are typically contaminated with a large number of white blood cells (WBCs). As a result, the diagnostic value of CTCs has been underused. Over the past decade, a diversity of CTC detection methods [2d, 3] have been developed to overcome the challenges encountered by the immunomagnetic-separation-based CellSearch assay. Different from the existing CTC diagnostic approaches,[2d, 3] we have demonstrated “nanovelcro” chips [4] that are capable of enriching, identifying, and enumerating CTCs in patient blood samples with superb efficiency. First, we pioneered a unique concept of nanovelcro substrates,[5] by which anti-EpCAM-coated silicon-nanowire (SiNW) substrates were used to immobilize CTCs in a stationary device. We have recently shown that other types of nanostructured substrates (eg, electrochemically deposited conjugated-polymer nanofeatures [6] and horizontally packed TiO2 nanofibers [7]) also exhibit enhanced affinity for capturing CTCs, demonstrating the general applicability of nanovelcro substrates. Our approach is unique because of the use of nanostructured substrates; there are enhanced local topographic interactions [8] between the anti-EpCAM-coated nanosubstrates and the nanoscaled cellular surface components (eg, microvilli) on a CTC, which are analogous to the working principle of velcro. Second, by integrating a lithographically patterned nanovelcro substrate with an overlaid poly (dimethylsiloxane)(PDMS) chaotic mixer [9] that enhances the frequency of contact between the CTCs flowing through the system and the substrate, we further improved CTC capture efficiency.[4] Side-by-side analytical validation studies using both artificial and patient CTC samples suggested that the sensitivity of nanovelcro chips outperformed [4] that of CellSearch. Although nanovelcro chips allow efficient and reproducible detection of CTCs in patient blood, challenges remain in 1) broadening its general applicability for detecting other types of solid-tumor CTCs that exhibit surface markers other than EpCAM, and 2) enabling the isolation of single CTCs for subsequent molecular analyses. To broaden the general applicability of the nanovelcro-based cell-affinity assay, we explored a melanoma-specific capture agent [13](ie, anti-CD146) to capture circulating melanoma cells (CMCs; a subcategory of solid-tumor CTCs). Further, an
登录
查看更多内容
影响因子：
3.2
作者：
Dickson, Mary Nora;Tsinberg, Pavel;Leonard, Edward F.
通讯作者：
Leonard, Edward F.
影响因子：
5.2
作者：
Rao, C.;Bui, T.;Terstappen, L. W. M. M.
通讯作者：
Terstappen, L. W. M. M.
DOI：
10.1111/j.1749-6632.2010.05779.x
发表时间：
2010-01-01
期刊：
TOWARD PERSONALIZED MEDICINE FOR CANCER
影响因子：
--
作者：
Riethdorf, Sabine;Pantel, Klaus
通讯作者：
Pantel, Klaus
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
11.5
作者：
Riethdorf, Sabine;Fritsche, Herbert;Pantel, Klaus
通讯作者：
Pantel, Klaus