Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology.
Label-free isolation of prostate circulating tumor cells using Vortex microfluidic technology.
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DOI：
10.1038/s41698-017-0015-0
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发表时间：
2017
影响因子：
7.9
通讯作者：
Sollier-Christen E
中科院分区：
文献类型：
--
作者：
Renier C;Pao E;Che J;Liu HE;Lemaire CA;Matsumoto M;Triboulet M;Srivinas S;Jeffrey SS;Rettig M;Kulkarni RP;Di Carlo D;Sollier-Christen E
人们越来越关注利用非侵入性“液体活检”来识别癌症预后和监测的生物标志物，以及分离可以预测靶向治疗反应的遗传物质。循环肿瘤细胞（CTC）已经成为这样一种生物标志物，提供有关肿瘤进化的遗传和表型信息，可能来自原发部位和转移部位。目前，可用的 CTC 分离方法，包括免疫亲和和基于尺寸的过滤，都侧重于高捕获效率，但纯度较低，并且通常需要较长的手动样品制备，这限制了捕获的 CTC 在下游分析中的使用。在这里，我们描述了使用微流控涡流芯片从 22 名晚期前列腺癌患者中进行基于大小的 CTC 分离，并且通过对其中 18 名患者的计数研究，发现我们可以在不到 1 小时内捕获高纯度（从 1.74 到 37.59%）和效率（从 1.88 到 93.75 CTC/7.5 mL）的 CTC。有趣的是，在五名年龄匹配的健康捐赠者（46-77 岁；1.25-2.50 CTC/7.5mL）中发现了更多的非典型大循环细胞，而在五名 <30 岁的健康捐赠者（21-27 岁；0.00 CTC/7.5mL）中发现了更多的非典型大循环细胞。使用根据 5 名年龄匹配的健康捐献者计算出的阈值 (3.37 CTC/mL)，我们在 80% 的前列腺癌患者中识别出了 CTC。我们还发现，收集的一部分细胞（11.5%）不表达上皮前列腺标记物（细胞角蛋白和/或前列腺特异性抗原），而有些细胞则表达上皮-间质转化标记物，即波形蛋白和 N-钙粘蛋白。我们还表明，分离细胞的纯度和 DNA 产量适合靶向扩增和新一代测序，无需全基因组扩增，可识别 15 个样本中的 10 个和 4 个健康样本中的 0 个中的独特突变。微流体装置可以快速有效地从前列腺癌患者的血液中分离循环肿瘤细胞。 Vortex Biosciences 的 Elodie Sollier-Christen、加州大学洛杉矶分校 David Geffen 医学院的 Rajan Kulkarni、加州大学洛杉矶分校的 Dino Di Carlo 及其同事对取自 21 名晚期前列腺癌男性和 10 名健康对照者的血液样本测试了该公司的微流体技术。他们表明，在一小时内，Vortex Chip 可以分离出 80% 癌症患者体内的循环肿瘤细胞，并且其中许多细胞不显示其他方法捕获前列腺癌细胞所需的常见表面标记。分离细胞的纯度和 DNA 产量足够高，可以进行靶向基因组测序，从而揭示可能与肿瘤形成有关的突变。 Vortex 技术可以帮助诊断前列腺癌并为患者的治疗决策提供信息。
There has been increased interest in utilizing non-invasive “liquid biopsies” to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on high capture efficiency but with lower purity and often long and manual sample preparation, which limits the use of captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can capture CTCs with high purity (from 1.74 to 37.59%) and efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46–77 years old; 1.25–2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21–27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial–mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples. A microfluidic device can rapidly and efficiently isolate circulating tumor cells from the blood of prostate cancer patients. Elodie Sollier-Christen of Vortex Biosciences, Rajan Kulkarni of David Geffen School of Medicine at UCLA, Dino Di Carlo of UCLA and colleagues tested the company’s microfluidic technology on blood samples taken from 21 men with advanced prostate cancer and 10 healthy controls. They showed that, within an hour, the Vortex Chip could isolate circulating tumor cells in 80% of the cancer patients and that many of these cells did not display the usual surface markers that other approaches require to capture prostate cancer cells. The purities and DNA yields of the isolated cells were high enough to enable targeted genome sequencing, which revealed mutations potentially involved in tumor formation. The Vortex technology could help diagnose prostate cancer and inform therapeutic decision-making for those with the disease.
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影响因子：
4.6
作者：
Dhar M;Pao E;Renier C;Go DE;Che J;Montoya R;Conrad R;Matsumoto M;Heirich K;Triboulet M;Rao J;Jeffrey SS;Garon EB;Goldman J;Rao NP;Kulkarni R;Sollier-Christen E;Di Carlo D
通讯作者：
Di Carlo D
影响因子：
9.2
作者：
Bitting, Rhonda L.;Schaeffer, Daneen;Somarelli, Jason A.;Garcia-Blanco, Mariano A.;Armstrong, Andrew J.
通讯作者：
Armstrong, Andrew J.
影响因子：
3.7
作者：
Dago AE;Stepansky A;Carlsson A;Luttgen M;Kendall J;Baslan T;Kolatkar A;Wigler M;Bethel K;Gross ME;Hicks J;Kuhn P
通讯作者：
Kuhn P
DOI：
10.1158/1078-0432.ccr-15-0137
发表时间：
2016-03-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Beltran H;Jendrisak A;Landers M;Mosquera JM;Kossai M;Louw J;Krupa R;Graf RP;Schreiber NA;Nanus DM;Tagawa ST;Marrinucci D;Dittamore R;Scher HI
通讯作者：
Scher HI
影响因子：
11.5
作者：
Danila, Daniel C.;Heller, Glenn;Scher, Howard I.
通讯作者：
Scher, Howard I.