Longitudinal cancer monitoring is crucial to clinical implementation of precision medicine. There is growing evidence indicating important functions of extracellular vesicles (EVs) in tumor progression and metastasis, including matrix remodeling via transporting matrix metalloproteases (MMPs). However, the clinical relevance of EVs remains largely undetermined, partially owing to challenges in EV analysis. Distinct from existing technologies mostly focused on characterizing molecular constituents of EVs, here we report a nanoengineered lab-on-a-chip system that enables integrative functional and molecular phenotyping of tumor-associated EVs. A generalized, high-resolution colloidal inkjet printing method was developed to allow robust and scalable manufacturing of three-dimensional (3D) nanopatterned devices. With this nanochip platform, we demonstrated integrative analysis of the expression and proteolytic activity of MMP14 on EVs to detect in vitro cell invasiveness and monitor in vivo tumor metastasis, using cancer cell lines and mouse models. Analysis of clinical plasma specimen showed that our technology could be used for cancer detection including accurate classification of age-matched controls and patients with ductal carcinoma in situ, invasive ductal carcinoma, or locally metastatic breast cancer in a training cohort (n = 30, 96.7% accuracy) and an independent validation cohort (n = 70, 92.9% accuracy). With clinical validation, our technology could provide a useful liquid biopsy tool to improve cancer diagnostics and real-time surveillance of tumor evolution in patients to inform personalized therapy.

癌症纵向监测对于精准医疗的临床实施至关重要。越来越多的证据表明细胞外囊泡 (EV) 在肿瘤进展和转移中发挥重要作用，包括通过运输基质金属蛋白酶 (MMP) 进行基质重塑。然而，EV 的临床相关性在很大程度上仍未确定，部分原因是 EV 分析面临挑战。与主要关注电动汽车分子成分特征的现有技术不同，我们在此报告了一种纳米工程芯片实验室系统，该系统能够对肿瘤相关电动汽车进行综合功能和分子表型分析。开发了一种通用的高分辨率胶体喷墨打印方法，可以稳健且可扩展地制造三维 (3D) 纳米图案器件。借助这个纳米芯片平台，我们利用癌细胞系和小鼠模型，对 EV 上 MMP14 的表达和蛋白水解活性进行了综合分析，以检测体外细胞侵袭性并监测体内肿瘤转移。对临床血浆样本的分析表明，我们的技术可用于癌症检测，包括对训练队列中年龄匹配的对照和导管原位癌、浸润性导管癌或局部转移性乳腺癌患者进行准确分类（ n = 30, 96.7） % 准确率）和独立验证队列（ n = 70，准确率 92.9%）。通过临床验证，我们的技术可以提供有用的液体活检工具，以改善癌症诊断和患者肿瘤演变的实时监测，从而为个性化治疗提供信息。