Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device.
Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device.
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DOI：
10.1371/journal.pone.0035976
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Giannakakou P
中科院分区：
文献类型：
--
作者：
Kirby BJ;Jodari M;Loftus MS;Gakhar G;Pratt ED;Chanel-Vos C;Gleghorn JP;Santana SM;Liu H;Smith JP;Navarro VN;Tagawa ST;Bander NH;Nanus DM;Giannakakou P
由于抗癌治疗反应不佳，癌症转移占癌症相关死亡的大部分。由于可用肿瘤组织的稀缺，对转移相关耐药性的分子理解仍然难以捉摸。从患者外周血中分离循环肿瘤细胞（CTC）已成为可进行分子表征的肿瘤组织的有效替代来源。然而，纯度和灵敏度低的问题阻碍了临床实践的采用。在这里，我们报告了一种新方法来捕获和分子表征从接受紫杉烷化疗的去势抵抗性前列腺癌患者（CRPC）中分离出的 CTC。我们开发了一种几何增强差分免疫捕获 (GEDI) 微流体装置，该装置将抗前列腺特异性膜抗原 (PSMA) 抗体与 3D 几何结构相结合，可捕获 CTC，同时最大限度地减少非特异性白细胞粘附。 GEDI 捕获的 CTC（定义为完整的有核 PSMA+/CD45− 细胞）计数显示，CRPC 患者中每毫升的中位数为 54 个细胞，而健康捐献者中为 3 个。与市售 CellSearch® 的直接比较显示，GEDI 设备的灵敏度提高了 2-400 倍。对患者来源的 GEDI 捕获的 CTC 进行共聚焦显微镜鉴定，发现了 TMPRSS2:ERG 融合蛋白，而测序则在仅掺有 50 个 PC C4-2 细胞的血液样本中发现了特定的雄激素受体点突变 (T868A)。使用多西他赛和紫杉醇对患者来源的 CTC 进行片上治疗，可以通过微管束来监测药物靶标的结合。从多西紫杉醇耐药的 CRPC 患者中分离出的 CTC 没有显示出任何药物活性的证据。这些测量构成了活体循环肿瘤细胞中药物靶标参与的第一个功能测定，因此有可能实现靶标反应的纵向监测并为新抗癌药物的开发提供信息。
Cancer metastasis accounts for the majority of cancer-related deaths owing to poor response to anticancer therapies. Molecular understanding of metastasis-associated drug resistance remains elusive due to the scarcity of available tumor tissue. Isolation of circulating tumor cells (CTCs) from the peripheral blood of patients has emerged as a valid alternative source of tumor tissue that can be subjected to molecular characterization. However, issues with low purity and sensitivity have impeded adoption to clinical practice. Here we report a novel method to capture and molecularly characterize CTCs isolated from castrate-resistant prostate cancer patients (CRPC) receiving taxane chemotherapy. We have developed a geometrically enhanced differential immunocapture (GEDI) microfluidic device that combines an anti-prostate specific membrane antigen (PSMA) antibody with a 3D geometry that captures CTCs while minimizing nonspecific leukocyte adhesion. Enumeration of GEDI-captured CTCs (defined as intact, nucleated PSMA+/CD45− cells) revealed a median of 54 cells per ml identified in CRPC patients versus 3 in healthy donors. Direct comparison with the commercially available CellSearch® revealed a 2–400 fold higher sensitivity achieved with the GEDI device. Confocal microscopy of patient-derived GEDI-captured CTCs identified the TMPRSS2:ERG fusion protein, while sequencing identified specific androgen receptor point mutation (T868A) in blood samples spiked with only 50 PC C4-2 cells. On-chip treatment of patient-derived CTCs with docetaxel and paclitaxel allowed monitoring of drug-target engagement by means of microtubule bundling. CTCs isolated from docetaxel-resistant CRPC patients did not show any evidence of drug activity. These measurements constitute the first functional assays of drug-target engagement in living circulating tumor cells and therefore have the potential to enable longitudinal monitoring of target response and inform the development of new anticancer agents.
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影响因子：
5.3
作者：
Gradilone A;Raimondi C;Nicolazzo C;Petracca A;Gandini O;Vincenzi B;Naso G;Aglianò AM;Cortesi E;Gazzaniga P
通讯作者：
Gazzaniga P
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
3.8
作者：
Gostner JM;Fong D;Wrulich OA;Lehne F;Zitt M;Hermann M;Krobitsch S;Martowicz A;Gastl G;Spizzo G
通讯作者：
Spizzo G
DOI：
10.1073/pnas.0404036101
发表时间：
2004-07-20
影响因子：
11.1
作者：
Krivacic, RT;Ladanyi, A;Bruce, RH
通讯作者：
Bruce, RH
影响因子：
11.5
作者：
Danila, Daniel C.;Heller, Glenn;Scher, Howard I.
通讯作者：
Scher, Howard I.