Integrated Analysis of Exosomal Protein Biomarkers on Alternating Current Electrokinetic Chips Enables Rapid Detection of Pancreatic Cancer in Patient Blood
Integrated Analysis of Exosomal Protein Biomarkers on Alternating Current Electrokinetic Chips Enables Rapid Detection of Pancreatic Cancer in Patient Blood
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DOI：
10.1021/acsnano.7b08199
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发表时间：
2018-04-01
期刊：
影响因子：
17.1
通讯作者：
Heller, Michael J.
中科院分区：
文献类型：
--
作者：
Lewis, Jean M.;Vyas, Ankit D.;Heller, Michael J.
胰腺导管腺癌（PDAC）通常具有非特异性症状，并且常常发现太晚而无法治疗。由于PDAC的诊断涉及复杂、侵入性和昂贵的程序，因此筛查风险增加的人群将依赖于开发快速、敏感、特异性和成本效益高的测试。外泌体是一种从肿瘤中脱落到血液中的纳米级囊泡，作为无创液体活检诊断的有价值实体而受到关注。然而，快速捕获和分析外泌体及其蛋白质和其他生物标志物已被证明是困难的。在这里，我们提出了一种简单的方法，将外泌体和其他细胞外囊泡直接从全血、血浆或血清中捕获和分析到交流电动微阵列芯片上。在此过程中，不需要预处理或稀释样品，也不需要使用捕获抗体或其他亲和技术。随后的片上免疫荧光分析允许在短短30分钟的总时间内对目标生物标志物进行特异性鉴定和定量。在这项初步验证研究中，发现生物标志物glypican-1和CD63反映了PDAC的存在，因此被用于建立检测PDAC的双变量模型。20例PDAC患者样本与11例健康受试者的灵敏度为99%，特异性为82%。在一小部分结肠癌患者样本中，观察到glypican-1在转移性疾病中升高，但在非转移性疾病中没有升高。ACE外泌体捕获和芯片上生物标志物检测的速度和简单性，加上使用全血的能力，将实现无缝的“从样本到答案”液体活检筛查，并改善早期癌症诊断。
Pancreatic ductal adenocarcinoma (PDAC) typically has nonspecific symptoms and is often found too late to treat. Because diagnosis of PDAC involves complex, invasive, and expensive procedures, screening populations at increased risk will depend on developing rapid, sensitive, specific, and cost-effective tests. Exosomes, which are nanoscale vesicles shed into blood from tumors, have come into focus as valuable entities for noninvasive liquid biopsy diagnostics. However, rapid capture and analysis of exosomes with their protein and other biomarkers have proven difficult. Here, we present a simple method integrating capture and analysis of exosomes and other extracellular vesicles directly from whole blood, plasma, or serum onto an AC electrokinetic microarray chip. In this process, no pretreatment or dilution of sample is required, nor is it necessary to use capture antibodies or other affinity techniques. Subsequent on-chip immunofluorescence analysis permits specific identification and quantification of target biomarkers within as little as 30 min total time. In this initial validation study, the biomarkers glypican-1 and CD63 were found to reflect the presence of PDAC and thus were used to develop a bivariate model for detecting PDAC. Twenty PDAC patient samples could be distinguished from 11 healthy subjects with 99% sensitivity and 82% specificity. In a smaller group of colon cancer patient samples, elevated glypican-1 was observed for metastatic but not for nonmetastatic disease. The speed and simplicity of ACE exosome capture and on-chip biomarker detection, combined with the ability to use whole blood, will enable seamless "sample-to-answer" liquid biopsy screening and improve early stage cancer diagnostics.