Molecular and functional extracellular vesicle analysis using nanopatterned microchips monitors tumor progression and metastasis

Abstract

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.

Dendrimer-decorated nanogels: Efficient nanocarriers for biodistribution in vivo and chemotherapy of ovarian carcinoma

“…Over the past few decades, the development of nanomedicine has revolutionized disease management via highly specific targeting or recognition, precision diagnosis, efficient treatment, and real-time monitoring [ [1] , [2] , [3] , [4] ]. However, the sophisticated biological environments in the body impose considerable barriers to the transport of cargoes for the targeted regions [ 5 , 6 ].…”

Microfluidic‐Based Exosome Analysis for Liquid Biopsy

“…reported a 3D‐nanopatterned microchip using a colloidal inkjet printing method. [ 117 ] They utilized this chip to analyze the expression and proteolytic activity of MMP14 on EVs using two small EV (sEV) enzyme‐linked immunosorbent assays. Although microfluidics‐based fluorescence technology has been applied for exosome detection with good accuracy and high sensitivity, shortcomings of this technology exist, such as the cumbersome preparation of fluorescent labels, spectral interferences.…”

“…applied nano‐HB to the molecular phenotyping of tumor‐related EVs. [ 117 ] By detecting EVs from clinical plasma specimens of the expression and activity of MMP14, the platform was able to accurately distinguish cancer patients from healthy controls of the same age and differentiate in situ ductal carcinoma, infiltrating ductal carcinoma and locally metastatic breast cancer. This platform could be used to improve the real‐time monitoring of tumor evolution or even guide personalized treatment by liquid biopsy.…”

The ins and outs of microvesicles

“…Researchers utilized microfluidic devices to examine phenotypic changes in melanoma extracellular vesicles to monitor treatment response, or to quantify the proteolytic activity of MT1-MMP for monitoring of in vivo tumor progression and metastasis. 79,80 As such, it is tempting to speculate on the future of EV clinical applications in exploiting the biochemical, physical, and functional makeup of EVs to improve diagnostics and real-time monitoring of disease.…”